Enhancing autophagy or increasing longevity by administration of urolithins

ABSTRACT

Disclosed are methods, compounds, and compositions useful for increasing autophagy and promoting longevity. The methods, compounds, and compositions relate to urolithins and urolithin precursors and use thereof. Certain urolithins are represented by Formula I, while certain urolithin precursors are represented by Formula IV. The urolithin may be urolithin A, urolithin B, urolithin C, or urolithin D. The urolithin precursor may be ellagic acid or an ellagitannin. The methods include in vivo, ex vivo, and in vitro uses of the compounds and compositions.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.17/324,490, filed May 19, 2021; which is a continuation of U.S. patentapplication Ser. No. 13/929,455, filed Jun. 27, 2013, now U.S. Pat. No.11,020,373; which claims the benefit of priority to U.S. ProvisionalPatent Application No. 61/791,137, filed Mar. 15, 2013; U.S. ProvisionalPatent Application No. 61/712,886, filed Oct. 12, 2012; and U.S.Provisional Patent Application No. 61/665,137, filed Jun. 27, 2012.

BACKGROUND

Autophagy is a lysosomal degradation pathway in both animals and plantsthat is essential for development, differentiation, homeostasis, andsurvival. In animals, autophagy serves principally as an adaptivemechanism to protect organisms against diverse pathologies, includinginfection, cancer, neurodegeneration, heart disease, and aging. Therepertoire of routine housekeeping functions performed by autophagyincludes elimination of defective proteins and organelles, prevention ofthe accumulation of abnormal protein aggregates, and elimination ofintracellular pathogens. The autophagy pathway is uniquely capable ofdegrading entire organelles, such as mitochondria, peroxisomes, andendoplasmic reticulum.

Multiple reports indicate that proteins required for autophagyinduction, such as sirtuin 1, have reduced expression in aged tissues;levels of autophagy have been shown to diminish with age. Reduced levelsof autophagy have also been associated with obesity, diabetes, cancer,neurodegenerative diseases, cardiovascular disease, osteoarthritis, andage-related macular degeneration.

A number of compounds that stimulate autophagy have been identified,including rapamycin, resveratrol, metformin, spermidine, andglucosamine.

Urolithins are ellagitannin- and ellagic acid-derived metabolitesproduced, e.g., by mammalian colonic microflora, including human colonicmicroflora. Urolithins are known to exhibit anti-oxidant activity.

SUMMARY OF THE INVENTION

An aspect of the invention is a method of increasing autophagy in ananimal, comprising the step of administering to an animal in needthereof an effective amount of a urolithin or a precursor thereof,thereby increasing autophagy in the animal.

An aspect of the invention is a method of increasing longevity in ananimal, comprising the step of administering to an animal in needthereof an effective amount of a urolithin or a precursor thereof,thereby increasing longevity of the animal.

An aspect of the invention is a method of increasing autophagy in acell, comprising the step of contacting a cell with an effective amountof a urolithin or a precursor thereof, thereby increasing autophagy inthe cell.

An aspect of the invention is a method of increasing longevity of acell, comprising the step of contacting a cell with an effective amountof a urolithin or a precursor thereof, thereby increasing longevity ofthe cell.

An aspect of the invention is a method of increasing autophagy ofeukaryotic cells in vitro, comprising the step of contacting eukaryoticcells in vitro with an effective amount of a urolithin or a precursorthereof, thereby increasing autophagy in the eukaryotic cells in vitro.

An aspect of the invention is a method of increasing longevity ofeukaryotic cells in vitro, comprising the step of contacting eukaryoticcells in vitro with an effective amount of a urolithin or a precursorthereof, thereby increasing longevity of the eukaryotic cells in vitro.

An aspect of the invention is a composition comprising a urolithin or aprecursor thereof; and a compound selected from the group consisting ofrapamycin, resveratrol, metformin, and spermidine.

An aspect of the invention is a compound of Formula II

wherein

X¹, X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are independently selected from thegroup consisting of H and OH; and

with the proviso that the compound is not a compound of Formula IIwherein

X¹, X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X¹ is OH, and X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X² is OH, and X¹, X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H (urolithin B);

X³ is OH, and X¹, X², X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X⁴ is OH, and X¹, X², X³, X⁵, X⁶, X⁷, and X⁸ are H;

X⁵ is OH, and X¹, X², X³, X⁴, X⁶, X⁷, and X⁸ are H;

X⁶ is OH, and X¹, X², X³, X⁴, X⁵, X⁷, and X⁸ are H;

X⁷ is OH, and X¹, X², X³, X⁴, X⁵, X⁶, and X⁸ are H;

X⁸ is OH, and X¹, X², X³, X⁴, X⁵, X⁶, and X⁷ are H;

X¹ and X² are OH, and X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X¹ and X⁵ are OH, and X², X³, X⁴, X⁶, X⁷, and X⁸ are H;

X¹ and X⁷ are OH, and X², X³, X⁴, X⁵, X⁶, and X⁸ are H;

X¹ and X⁸ are OH, and X², X³, X⁴, X⁵, X⁶, and X⁷ are H;

X² and X³ are OH, and X¹, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X² and X⁴ are OH, and X¹, X³, X⁵, X⁶, X⁷, and X⁸ are H;

X² and X⁵ are OH, and X¹, X³, X⁴, X⁶, X⁷, and X⁸ are H;

X² and X⁶ are OH, and X¹, X³, X⁴, X⁵, X⁷, and X⁸ are H (urolithin A);

X² and X⁷ are OH, and X¹, X³, X⁴, X⁵, X⁶, and X⁸ are H;

X³ and X⁴ are OH, and X¹, X², X⁵, X⁶, X⁷, and X⁸ are H;

X³ and X⁵ are OH, and X¹, X², X⁴, X⁶, X⁷, and X⁸ are H;

X³ and X⁶ are OH, and X¹, X², X⁴, X⁵, X⁷, and X⁸ are H;

X⁵ and X⁶ are OH, and X¹, X², X³, X⁴, X⁷, and X⁸ are H;

X⁵ and X⁸ are OH, and X¹, X², X³, X⁴, X⁶, and X⁷ are H;

X⁶ and X⁷ are OH, and X¹, X², X³, X⁴, X⁵, and X⁸ are H;

X¹, X², and X⁵ are OH, and X³, X⁴, X⁶, X⁷, and X⁸ are H;

X¹, X², and X⁶ are OH, and X³, X⁴, X⁵, X⁷, and X⁸ are H;

X¹, X⁵, and X⁸ are OH, and X², X³, X⁴, X⁶, and X⁷ are H;

X², X⁴, and X⁶ are OH, and X¹, X³, X⁵, X⁷, and X⁸ are H;

X², X⁴, and X⁷ are OH, and X¹, X³, X⁵, X⁶, and X⁸ are H;

X², X⁶, and X⁷ are OH, and X¹, X³, X⁴, X⁵, and X⁸ are H (urolithin C);

X², X⁶, and X⁸ are OH, and X¹, X³, X⁴, X⁵, and X⁷ are H;

X², X⁷, and X⁸ are OH, and X¹, X³, X⁴, X⁵, and X⁶ are H;

X¹, X², X⁵, and X⁶ are OH, and X³, X⁴, X⁷, and X⁸ are H;

X¹, X², X⁵, and X⁷ are OH, and X³, X⁴, X⁶, and X⁸ are H;

X¹, X², X⁶, and X⁷ are OH, and X³, X⁴, X⁵, and X⁸ are H (urolithin D);

X¹, X⁶, X⁷, and X⁸ are OH, and X², X³, X⁴, and X⁵ are H;

X², X³, X⁶, and X⁷ are OH, and X¹, X⁴, X⁵, and X⁸ are H;

X², X⁴, X⁵, and X⁸ are OH, and X¹, X³, X⁶, and X⁷ are H;

X², X⁴, X⁶, and X⁷ are OH, and X¹, X³, X⁵, and X⁸ are H;

X¹, X², X⁴, X⁵, and X⁷ are OH, and X³, X⁶, and X⁸ are H;

X¹, X², X⁶, X⁷, and X⁸ are OH, and X³, X⁴, and X⁵ are H; and

X¹, X², X³, X⁶, X⁷, and X⁸ are OH, and X⁴ and X⁵ are H.

An aspect of the invention is a compound of Formula III

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR;

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide; and

with the proviso that the compound is not a compound of Formula IIIwherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R¹ is OR, and R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R² is OR, and R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R³ is OR, and R¹, R², R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R⁴ is OR, and R¹, R², R³, R⁵, R⁶, R⁷, and R⁸ are H;

R⁵ is OR, and R¹, R², R³, R⁴, R⁶, R⁷, and R⁸ are H;

R⁶ is OR, and R¹, R², R³, R⁴, R⁵, R⁷, and R⁸ are H;

R⁷ is OR, and R¹, R², R³, R⁴, R⁵, R⁶, and R⁸ are H;

R⁸ is OR, and R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are H;

R¹ and R² are OR, and R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R¹ and R⁵ are OR, and R², R³, R⁴, R⁶, R⁷, and R⁸ are H;

R¹ and R⁷ are OR, and R², R³, R⁴, R⁵, R⁶, and R⁸ are H;

R¹ and R⁸ are OR, and R², R³, R⁴, R⁵, R⁶, and R⁷ are H;

R² and R³ are OR, and R¹, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R² and R⁴ are OR, and R¹, R³, R⁵, R⁶, R⁷, and R⁸ are H;

R² and R⁵ are OR, and R¹, R³, R⁴, R⁶, R⁷, and R⁸ are H;

R² and R⁶ are OR, and R¹, R³, R⁴, R⁵, R⁷, and R⁸ are H;

R² and R⁷ are OR, and R¹, R³, R⁴, R⁵, R⁶, and R⁸ are H;

R² and R⁸ are OR, and R¹, R³, R⁴, R⁵, R⁶, and R⁷ are H;

R³ and R⁴ are OR, and R¹, R², R⁵, R⁶, R⁷, and R⁸ are H;

R³ and R⁵ are OR, and R¹, R², R⁴, R⁶, R⁷, and R⁸ are H;

R³ and R⁶ are OR, and R¹, R², R⁴, R⁵, R⁷, and R⁸ are H;

R³ and R⁷ are OR, and R¹, R², R⁴, R⁵, R⁶, and R⁸ are H;

R³ and R⁸ are OR, and R¹, R², R⁴, R⁵, R⁶, and R⁷ are H;

R⁴ and R⁸ are OR, and R¹, R², R³, R⁵, R⁶, and R⁷ are H;

R⁵ and R⁶ are OR, and R¹, R², R³, R⁴, R⁷, and R⁸ are H;

R⁵ and R⁷ are OR, and R¹, R², R³, R⁴, R⁶, and R⁸ are H;

R⁵ and R⁸ are OR, and R¹, R², R³, R⁴, R⁶, and R⁷ are H;

R⁶ and R⁷ are OR, and R¹, R², R³, R⁴, R⁵, and R⁸ are H;

R⁶ and R⁸ are OR, and R¹, R², R³, R⁴, R⁵, and R⁷ are H;

R¹, R², and R³ are OR, and R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R¹, R², and R⁵ are OR, and R³, R⁴, R⁶, R⁷, and R⁸ are H;

R¹, R², and R⁶ are OR, and R³, R⁴, R⁵, R⁷, and R⁸ are H;

R¹, R², and R⁸ are OR, and R³, R⁴, R⁵, R⁶, and R⁷ are H;

R¹, R⁵, and R⁸ are OR, and R², R³, R⁴, R⁶, and R⁷ are H;

R¹, R⁷, and R⁸ are OR, and R², R³, R⁴, R⁵, and R⁶ are H;

R², R³, and R⁴ are OR, and R¹, R⁵, R⁶, R⁷, and R⁸ are H;

R², R⁴, and R⁶ are OR, and R¹, R³, R⁵, R⁷, and R⁸ are H;

R², R⁴, and R⁷ are OR, and R¹, R³, R⁵, R⁶, and R⁸ are H;

R², R⁵, and R⁸ are OR, and R¹, R³, R⁴, R⁶, and R⁷ are H;

R², R⁶, and R⁷ are OR, and R¹, R³, R⁴, R⁵, and R⁸ are H;

R², R⁶, and R⁸ are OR, and R¹, R³, R⁴, R⁵, and R⁷ are H;

R², R⁷, and R⁸ are OR, and R¹, R³, R⁴, R⁵, and R⁶ are H;

R³, R⁵, and R⁸ are OR, and R¹, R², R⁴, R⁶, and R⁷ are H;

R³, R⁷, and R⁸ are OR, and R¹, R², R⁴, R⁵, and R⁶ are H;

R⁶, R⁷, and R⁸ are OR, and R¹, R², R³, R⁴, and R⁵ are H;

R¹, R², R⁵, and R⁶ are OR, and R³, R⁴, R⁷, and R⁸ are H;

R¹, R², R⁵, and R⁷ are OR, and R³, R⁴, R⁶, and R⁸ are H;

R¹, R², R⁶, and R⁷ are OR, and R³, R⁴, R⁵, and R⁸ are H;

R¹, R⁶, R⁷, and R⁸ are OR, and R², R³, R⁴, and R⁵ are H;

R², R³, R⁴, and R⁶ are OR, and R¹, R⁵, R⁷, and R⁸ are H;

R², R³, R⁵, and R⁷ are OR, and R¹, R⁴, R⁶, and R⁸ are H;

R², R³, R⁶, and R⁷ are OR, and R¹, R⁴, R⁵, and R⁸ are H;

R², R⁴, R⁵, and R⁸ are OR, and R¹, R³, R⁶, and R⁷ are H;

R², R⁴, R⁶, and R⁷ are OR, and R¹, R³, R⁵, and R⁸ are H;

R², R⁵, R⁶, and R⁷ are OR, and R¹, R³, R⁴, and R⁸ are H;

R², R⁶, R⁷, and R⁸ are OR, and R¹, R³, R⁴, and R⁵ are H;

R¹, R², R⁴, R⁵, and R⁷ are OR, and R³, R⁶, and R⁸ are H;

R¹, R², R⁶, R⁷, and R⁸ are OR, and R³, R⁴, and R⁵ are H;

R², R³, R⁴, R⁵, and R⁷ are OR, and R¹, R⁶, and R⁸ are H;

R², R³, R⁶, R⁷, and R⁸ are OR, and R¹, R⁴, and R⁵ are H;

R², R⁴, R⁶, R⁷, and R⁸ are OR, and R¹, R³, and R⁵ are H;

R², R⁵, R⁶, R⁷, and R⁸ are OR, and R¹, R³, and R⁴ are H;

R¹, R², R³, R⁶, R⁷, and R⁸ are OR, and R⁴ and R⁵ are H;

R², R³, R⁴, R⁶, R⁷, and R⁸ are OR, and R¹ and R⁵ are H; and

R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR, and R¹ and R³ are H.

An aspect of the invention is a compound of Formula V

wherein

X⁹, X¹⁰, X¹¹, X¹², X¹³, and X¹⁴ are independently selected from thegroup consisting of H and OH; and with the proviso that the compound isnot a compound of Formula V wherein

X⁹, X¹⁰, X¹¹, X¹², X¹³, and X¹⁴ are H;

X¹⁰ is OH, and X⁹, X¹¹, X¹², X¹³, and X¹⁴ are H;

X⁹ and X¹² are OH, and X¹⁰, X¹¹, X¹³, and X¹⁴ are H;

X⁹ and X¹³ are OH, and X¹⁰, X¹¹, X¹², and X¹⁴ are H;

X⁹ and X¹⁴ are OH, and X¹⁰, X¹¹, X¹², and X¹³ are H;

X¹⁰ and X¹³ are OH, and X⁹, X¹¹, X¹², and X¹⁴ are H;

X¹⁰, X¹¹, and X¹³ are OH, and X⁹, X¹², and X¹⁴ are H;

X⁹, X¹⁰, X¹², and X¹⁴ are OH, and X¹¹ and X¹³ are H;

X⁹, X¹⁰, X¹³, and X¹⁴ are OH, and X¹¹ and X¹² are H (ellagic acid);

X⁹, X¹⁰, X¹¹, X¹³ and X¹⁴ are OH, and X¹² is H; and

X⁹, X¹⁰, X¹¹, X¹², X¹³, and X¹⁴ are OH.

An aspect of the invention is a compound of Formula VI

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR;

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide; and

with the proviso that the compound is not a compound of Formula VIwherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are H;

R¹⁰ is OR, and R⁹, R¹¹, R¹², R¹³, and R¹⁴ are H;

R⁹ and R¹² are OR, and R¹⁰, R¹¹, R¹³, and R¹⁴ are H;

R⁹ and R¹³ are OR, and R¹⁰, R¹¹, R¹², and R¹⁴ are H;

R⁹ and R¹⁴ are OR, and R¹⁰, R¹¹, R¹², and R¹³ are H;

R¹⁰ and R¹³ are OR, and R⁹, R¹¹, R¹², and R¹⁴ are H;

R⁹, R¹⁰, and R¹³ are OR, and R¹¹, R¹², and R¹⁴ are H;

R⁹, R¹⁰, and R¹⁴ are OR, and R¹¹, R¹², and R¹³ are H;

R¹⁰, R¹¹, and R¹³ are OR, and R⁹, R¹², and R¹⁴ are H;

R⁹, R¹⁰, R¹², and R¹³ are OR, and R¹¹ and R¹⁴ are H;

R⁹, R¹⁰, R¹², and R¹⁴ are OR, and R¹¹ and R¹³ are H;

R⁹, R¹⁰, R¹³, and R¹⁴ are OR, and R¹¹ and R¹² are H;

R¹⁰, R¹¹, R¹², and R¹³ are OR, and R⁹ and R¹⁴ are H;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ are OR, and R¹⁴ is H;

R⁹, R¹⁰, R¹¹, R¹³, and R¹⁴ are OR, and R¹² is H; and

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are OR.

An aspect of the invention is a composition comprising a first compound;and a second compound selected from the group consisting of rapamycin,resveratrol, metformin, and spermidine, wherein the first compound is acompound of any one of Formulas II, III, V, or VI.

An aspect of the invention is a method of increasing autophagy in acell, comprising contacting the cell with an effective amount of acompound of any one of Formulas II, III, V, or VI, thereby increasingautophagy in the cell.

An aspect of the invention is a method of increasing longevity in ananimal, comprising administering to an animal in need thereof aneffective amount of a compound of any one of Formulas II, III, V, or VI,thereby increasing longevity of the animal.

An aspect of the invention is a method of increasing longevity ofeukaryotic cells in vitro, comprising contacting eukaryotic cells invitro with an effective amount of a compound of any one of Formulas II,III, V, or VI, thereby increasing longevity of the eukaryotic cells invitro.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic diagram depicting four steps of macroautophagy:induction and nucleation, expansion, fusion, and degradation. Proteinsinvolved in each step are indicated above each step. The role of p62 andLC3 is explained schematically, whereby p62 helps to transport cellularmaterial into the autophagosome by binding to LC3.

FIG. 2 depicts structural formulas for urolithin A (UA), ellagic acid(EA), tellimagrandin (TL), punicalagin (PA), and punicalin (PB).

FIG. 3 depicts ellagic acid (EA) and its metabolites, urolithin D (UD),urolithin C (UC), urolithin A (UA), and urolithin B (UB), which areproduced by intestinal microflora in mammals, including humans.

FIG. 4 is a group of five graphs depicting the effect of ellagic acidand urolithins A, B, C, and D on the lifespan of C. elegans. Test agentswere present at 50 μM in DMSO. DMSO, dimethylsulfoxide, was the controland vehicle for test agents.

FIG. 5 is a set of graphs depicting longevity of wild-type C. elegansgrown in the presence of urolithin A at the concentrations shown.

FIGS. 6A-6F are a group of six graphs depicting lifespan analysis ofwild-type and indicated mutant strains of C. elegans grown in theabsence (black) or presence (grey) of urolithin A at 50 μM.

FIG. 7 is a bar graph depicting the effect of urolithin A onmitochondria in muscle of C. elegans. Transgenic C. elegans strainSJ4103 shows fluorescence due to muscle-specific expression of greenfluorescent protein (GFP) which is targeted to the mitochondrialmembrane. Mitochondria presence in the muscle of the C. elegans is shownby an increase in fluorescence. Results are expressed as mean±SEM. *,p=0.0014 (Student's t-test).

FIGS. 8A-8D are a line graph and three bar graphs depicting effect ofurolithin A (UA) on basal and uncoupled respiration in young(one-day-old) and old (ten-day-old) C. elegans. FIG. 8A illustratesbasal and uncoupled respiration (FCCP) in 10-day-old control wormstreated with 0.1% DMSO and ten-day-old-worms treated with 30 μMurolithin A in 0.1% DMSO. FIG. 8B illustrates representative area underthe curve (AUC) of uncoupled (FCCP) respiration in ten-day-old controlworms treated with vehicle (0.1% DMSO) or 30 μM urolithin A in 0.1%DMSO. Results are expressed as mean±SEM. *, p<0.05 (Student's t-test).OCR, oxygen consumption rate. FIG. 8C illustrates comparison of basalrespiration between one-day-old and ten-day-old worms treated withvehicle (0.1% DMSO). FIG. 8D illustrates comparison of basal respirationbetween one-day-old and ten-day-old worms treated with UA (30 μM).

FIG. 9A is a group of three confocal images depicting the effect ofurolithin A on autophagy induction in C. elegans.

FIG. 9B is a corresponding dot graph depicting the effect of urolithin Aon autophagy induction. ***, p<0.001 (student's t-test).

FIGS. 10A-10C are three graphs depicting survival curves, showing theeffect of inactivation by RNAi of vps-34 and bec-1 on the longevityphenotype induced by urolithin A treatment in C. elegans. Both vps-34(FIG. 10B) and bec-1 (FIG. 10C) inhibitions totally suppress thelifespan phenotype observed in worms treated with urolithin A (50 μM)and fed with empty vector (FIG. 10A). Survival analyses were performedusing the Kaplan Meier method and the significance of differencesbetween survival curves calculated using the log rank test. ***, p<0.001(log rank test).

FIG. 11 is a set of graphs demonstrating the effects of urolithin A(UA), urolithin B (UB), urolithin C (UC) and urolithin D (UD) treatmenton pharyngeal pumping in C. elegans worms after 7 and 14 days oftreatment. (*p<0.05; **p<0.01; ***p<0.001)

FIG. 12 are three line graphs demonstrating the effects of ellagic acid(EA), urolithin A (UA) and urolithin B (UB) treatment on motility inyoung C. elegans worms at days 1, 3, 5, and 8 of treatment. (*p<0.05;**p<0.01; ***p<0.001)

FIG. 13 is a set of images demonstrating time-lapse traces of C. elegansmotility following treatment with ellagic acid (EA), urolithin A (UA),urolithin B (UB), urolithin C (UC) and urolithin D (UD) treatment ondays 8, 14 and 16 of treatment.

FIG. 14 is a western blot of ModeK cells, a mouse intestinal epithelialcell line, demonstrating the effect of urolithin A treatment on theautophagy marker ratio LC3-II/LC3-I, p62, and on the ratio ofp-AMPKα/AMPKα. Bar graph demonstrates the quantified fold increase inthe ratio of LC3-II to LC3-I, and the ratio of p-AMPKα to AMPKα levelsobserved in the western blots. ctl, control.

FIG. 15 is a western blot of primary mouse hepatocytes, demonstratingthe effect of urolithin A treatment on the autophagy marker ratioLC3-II/LC3-I, p62, and on the ratio of p-AMPKα/AMPKα. Bar graphdemonstrates the quantified fold increase in the ratio of LC3-II toLC3-I, and the ratio of p-AMPKα to AMPKα levels observed in the westernblots. ctl, control.

FIG. 16 is a western blot of C2Cl2 mouse myocytes, demonstrating theeffect of urolithin A treatment on the autophagy marker ratioLC3-II/LC3-I, p62, and on the ratio of p-AMPKα/AMPKα. Bar graphdemonstrates the quantified fold increase in the ratio of LC3-II toLC3-I, and the ratio of p-AMPKα to AMPKα levels observed in the westernblots. ctl, control.

FIG. 17 is a western blot of human primary myoblasts, demonstrating theeffect of urolithin A treatment on the autophagy marker ratioLC3-II/LC3-I, p62, and on the ratio of p-AMPKα/AMPKα. Bar graphdemonstrates the quantified fold increase in the ratio of LC3-II toLC3-I levels observed in the western blots. Ctrl, control.

FIG. 18 is a western blot of human primary aortic endothelial cells,demonstrating the effect of urolithin A treatment on the autophagymarker ratio LC3-II/LC3-I and the protein p62. Bar graph demonstratesthe quantified fold increase in the ratio of LC3-II to LC3-I levelsobserved in the western blots. Ctrl, control.

FIG. 19 is a western blot of livers isolated from untreated control miceand mice administered urolithin A at a dose of 55 mg/kg/day admixed infood. Urolithin A treatment increased the autophagy marker ratioLC3-II/LC3-I, decreased p62, and increased the ratio of p-AMPKα/AMPKα.Bar graph demonstrates the quantified fold increase in the ratio ofLC3-II to LC3-I, and the ratio of p-AMPKα to AMPKα levels observed inthe western blots. ctl, control.

FIG. 20 is a graph depicting the effect of orally consumed urolithin Aat 55 mg/kg/day on the motor activity of C57BL/6J mice. Young treatedmice increased their spontaneous voluntary running on a running wheel byat least 25% during the five-day period investigated.

FIG. 21 is a bar graph depicting the effect of orally consumed urolithinA on running in aged C57BL/6J mice.

FIG. 22 is a bar graph depicting the effect of orally consumed urolithinA on grip strength in aged C57BL/6J mice.

FIG. 23 is pair of graphs depicting the effect of orally consumedurolithin A on ambulation and rearing in aged C57BL/6J mice. HFD, highfat diet; UA, urolithin A.

FIG. 24 is a western blot of skeletal muscle isolated from aged high fatdiet (HFD) untreated control mice, and from aged high fat diet miceadministered urolithin A (UA) at a dose of 50 mg/kg/day admixed in food.Urolithin A treatment increased the autophagy marker ratio LC3-II/LC3-Iand decreased the levels of p62. Bar graph demonstrates the quantifiedfold increase in the ratio of LC3-II to LC3-I levels observed in thewestern blots. Ctrl, control.

FIG. 25 shows the effect of urolithin A (UA) on autophagy in C2Cl2myoblasts. Myoblasts incubated 24 hrs with increasing doses of UA showeda dose response, with increasing autophagy as the UA concentration wasraised (10 μM, 50 μM and 100 μM), which was demonstrated by theincreasing shift in the histogram representing the LC3-B cell levels, amarker of autophagy, as compared to untreated controls.

FIG. 26 shows the effect of urolithin A (UA), urolithin B (UB),urolithin C (UC), and urolithin D (UD) on autophagy in C2Cl2 cells.Myoblasts incubated with UA, UB, UC, or UD at 100 μM experienced anincrease in autophagy as demonstrated by the shift in the histogramrepresenting the LC3-B cell levels as compared to untreated controls.

FIG. 27 depicts twenty-five compounds of the invention.

FIG. 28 depicts prophetic synthetic routes to the compounds in FIG. 27 .

DETAILED DESCRIPTION OF THE INVENTION Overview

Autophagy is a process by which cells degrade their own components,recycling amino acids and other building blocks that can be reused. Suchdegradation is performed by lysosomal acidic hydrolases. It is a tightlyregulated process that plays an important role in normal cell growth,development, and homeostasis, helping to maintain a balance between thesynthesis, degradation, and subsequent recycling of cellular products.It is a major mechanism by which starving cells can reallocate nutrientsfrom less-essential processes to more essential processes.

During nutrient starvation, increased levels of autophagy lead to thebreakdown of non-vital components and the release of nutrients, ensuringthat vital processes can continue. Mutant yeast cells that have areduced autophagic capability rapidly perish in nutrient-deficientconditions. A gene known as Atg7 has been implicated innutrient-mediated autophagy, and studies in mice have shown thatstarvation-induced autophagy was impaired in Atg7-deficient mice.Komatsu M et al. (2005) J Cell Biol. 169:425-434.

Autophagy degrades damaged organelles, cell membranes, and proteins. Thefailure of autophagy is thought to be an important factor in theaccumulation of cell damage and, therefore, aging.

Three types of autophagy can be distinguished, depending on the pathwayalong which cellular components are delivered to lysosomes:macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA).

Macroautophagy

Macroautophagy involves the degradation of long-lived proteins and wholecellular organelles through a multistep process (FIG. 1 ).Macroautophagy begins with the formation of a double-layered isolationmembrane (phagophore) around the molecules and/or organelles to bedegraded. The phagophore engulfs cytosolic components and seals aroundthe content, forming an autophagosome. Eventually, the autophagosomefuses with a lysosome, evolving into an autophagolysosome (orautolysosome), wherein lysosomal hydrolases digest the cargo.Microautophagy involves the direct sequestration of cytosolic componentsthrough invaginations or armlike projections of the lysosomal membrane.Microautophagy may serve for the turnover of long-lived proteins;however, the significance and regulation of this type of autophagyremain poorly understood. Finally, chaperone-mediated autophagy is ahighly selective process devoted to the degradation of soluble cytosolicproteins.

The microtubule-associated protein 1A/1B-light chain 3 (LC3), amammalian homolog of the yeast Atg8, is a soluble protein with amolecular mass of approximately 17 kDa which is distributed ubiquitouslyin mammalian tissues and cultured cells. It is processed immediatelyafter its synthesis by Atg4B, a cysteine protease, that exposes theC-terminal glycine residue (LC3-I). During autophagy, autophagosomesengulf cytoplasmic components, including cytosolic proteins andorganelles. Concomitantly, a cytosolic form of LC3 (LC3-I) is conjugatedto phosphatidylethanolamine (PE) to form and LC3-PE conjugate (LC3-II),which is recruited to autophagosomal membranes (FIG. 1 ).

p62, also known as sequestosome-1, was identified as a novel partner ofthe atypical protein kinase Cs (aPKCs) and is a ubiquitiously expressedcellular protein. p62 is known to have domains that interact with andbind to ubiquitinated proteins, and it has been identified as acomponent of inclusion bodies observed in human diseases, especiallyneurodegenerative diseases (e.g., Alzheimer's disease, Parkinson'sdisease, amyotrophic lateral sclerosis) as well as in liver diseases.p62 also has been identified as an LC3 interacting protein and it hasbeen demonstrated that an 11 amino acid sequence in the mouse p62 servesto recognize the LC3 protein. As seen in FIG. 1 , LC3 binds to p62 andtransports it (and any ubiquitinated proteins or cell components boundto it) into the autophagosome, where it is degraded. Consequently, oneof the hallmarks of autophagy is an increase in the ratio of LC3-II/LC34with a concomitant decrease in the level of cellular p62.

Of the three types of autophagy described, macroautophagy is the bestcharacterized in mammalian cells. Starvation is the strongest stimulusof macroautophagy. During nutrient deprivation, macroautophagy breaksdown cellular components, generating amino acids, fatty acids, andcarbohydrates, which can be harnessed for energy production and for thesynthesis of essential cellular molecules. Macroautophagy is alsoinvolved in specific cytosolic rearrangements during embryogenesis andpostnatal development. Furthermore, macroautophagy is induced duringviral or bacterial infections, in hypoxia, and under various stressconditions, including radiation exposure and increased reactive oxygenspecies (ROS) generation. In these circumstances, macroautophagy isessential for the maintenance of cell homeostasis by its promotion ofthe removal of damaged components. Indeed, impairments in macroautophagyinduce premature aging and shorten the lifespan in several organisms,including C. elegans, yeast, and Drosophila. Hars E S et al. (2007)Autophagy 3:93-95; Matecic M et al. (2010) PLoS Genet. 6:e1000921; Lee JH et al. (2010) Science 327:1223-1228. Conversely, upregulation ofmacroautophagy is proposed to be a major mechanism underlying thelifespan-extending properties of calorie restriction. Toth M L et al.(2008) Autophagy 4:330-338; Morselli E et al. (2010) Cell Death Dis.1:e10.

More than 35 Atg (AuTophaGy-related) proteins have been identified inyeasts and mammals; however, the precise role each Atg protein playsduring autophagy is not yet fully established. As illustrated in FIG. 1, the process of macroautophagy can be divided into discrete steps,namely, induction and nucleation, expansion, fusion, and degradation.The induction phase is mediated by the ULK1-Atg13-FIP200 kinase complex.The regulation of the nucleation stage, which consists of therecruitment of Atg proteins to the phagophore assembly site, is not yetcompletely understood. However, the vacuolar protein sorting-34 (Vps34),a class III phosphatidylinositol-3-kinase (PI3K), is required for thisstep. Vps34 associates with Beclin1, the mammalian homologue of yeastAtg6, and subsequently recruits Atg14 and Vps15 (p150) to thepreautophagosomal structure. The elongation and expansion of thephagophore membrane require two ubiquitin-like conjugation systemsinvolving Atg12 (conjugated to Atg5) and Atg8/microtubule-associatedprotein 1 light chain-3 (LC3, conjugated to phosphatidyl ethanolamine),along with other Atg proteins such as Atg9 and Atg16. The fusion of theautophagosome with a lysosome relies on canonical cellular fusionmachinery that consists of the Rab-SNARE (SolubleN-ethylmaleimide-sensitive factor Attachment protein REceptor) systemand requires the presence of lysosomal membrane-associated protein-2(LAMP-2) and the UV radiation resistance-associated gene (UVRAG).Finally, the digestion of the cargo is accomplished by lysosomalhydrolases, followed by the transportation of degraded components intothe cytoplasm by lysosomal efflux transporters such as Atg22.

With regard to the regulation of macroautophagy, mTOR, the mammaliantarget of rapamycin, is considered to be a major checkpoint, linking thecellular nutritional state with the level of ongoing autophagy. Undernutrient-rich conditions, mTOR is active and inhibits theULK1-Atg13-FIP200 complex required for the induction of macroautophagy.Energy deprivation leads to mTOR inactivation and stimulation ofAMP-activated protein kinase (AMPK), which both induce macroautophagy.AMPK functions as an energy-sensing kinase and is activated by increasesin the cellular AMP to ATP ratio. Under such circumstances, AMPKpromotes autophagy by directly activating ULK1 and by relieving themTOR-mediated inhibition of macroautophagy.

Macroautophagy can be selectively directed toward the removal ofparticular targets, e.g., peroxisomes (pexophagy), endoplasmic reticulum(reticulophagy), intracellular lipids (lipophagy), ribosomes(ribophagy), and intracellular pathogens (xenopathy). Likewise,mitochondria can be selectively targeted for degradation viamacroautophagy (mitophagy).

Mitophagy: A Specialized Form of Macroautophagy

Mitophagy is a highly selective process that can promote the eliminationof dysfunctional or unnecessary mitochondria. Wang K et al. (2011)Autophagy 7:297-300. The loss of mitochondrial membrane potential(Δψ_(m)) represents a major trigger of mitophagy. Indeed, laser-inducedphoto damage of selected mitochondria inside living hepatocytes resultsin the rapid dissipation of Δψ_(m), followed by the quick removal ofdepolarized mitochondria through mitophagy. In addition, oxidativedamage can lead to the formation of asymmetrical daughter mitochondriacharacterized by different Δψ_(m), with autophagy specifically targetingmitochondria with lower Δψ_(m). Apart from the degradation of damagedmitochondria under stress conditions, mitophagy is essential formitochondrial turnover in the basal state and during celldifferentiation, such as the maturation of reticulocytes into mature redblood cells.

Investigations into the molecular regulation of mitophagy have unveiledseveral mitophagy-specific proteins. Parkin and Pink1 are believed toplay important roles in the selective degradation of damagedmitochondria, at least under certain circumstances. Parkin is acytosolic E3-ubiquitin ligase that is selectively recruited todysfunctional mitochondria and assists in their removal by mitophagy.Narenda D (2008) J Cell Biol. 183:795-803. Pink1 is imported intohealthy mitochondria through a Δψ_(m)-dependent process and is degradedby the presenilin-associated rhomboidlike (PARL) protease. Matsuda N etal. (2010) J Cell Biol. 189:211-221. The dissipation of Δψ_(m) resultsin the accumulation of Pink1 on the mitochondrial surface, leading tothe recruitment of Parkin, which ubiquitinates outer membrane proteins,including the voltage-dependent anion channel (VDAC). It is proposedthat ubiquitin-tagged mitochondria are targeted directly to autophagicvacuoles through the interaction of ubiquitinated proteins with theautophagosomal marker LC3 (Atg8). In addition, Parkin can ubiquitinatethe inner mitochondrial membrane and apoptosis regulator protein B-celllymphoma-2 (Bcl-2), thereby de-repressing Beclin1.

Recent evidence also suggests that the opening of the mitochondrialpermeability transition pore (mPTP) may be required for the selectiveremoval of damaged mitochondria. Opening of the mPTP causes a suddenincrease of the inner membrane permeability to solutes with molecularweight up to 1500 Da. This results in mitochondrial depolarization,activation of the mitochondrial ATPase (i.e., ATP synthase operating inreverse), and swelling and rupture of the outer membrane. The loss ofΔψ_(n), subsequent to permeability transition targets individualmitochondria for degradation. The loss of Δψ_(n), and the activation ofmacroautophagy are prevented by cyclosporin A, an inhibitor of the mPTPcomponent cyclophilin D. Furthermore, starvation fails to inducemacroautophagy in cyclophilin D-deficient murine cardiomyocytes, whereasautophagy is enhanced even under fed conditions in cardiac cells frommice overexpressing cyclophilin D. The nicotinamide adenine dinucleotide(NAD)-dependent deacetylase sirtuin-3 (SIRT3) appears to be criticallyinvolved in the control of mPTP by modulation of cyclophilin D.

Similar to the mPTP, the apoptotic proteins Bnip3 (Bcl-2 and adenovirusE1B 19-kDa-interacting protein-3) and Nix (Nip3-like protein X) arethought to trigger selective mitophagy through mitochondrialdepolarization. Moreover, Bnip3 may induce mitophagy by competitivelydisrupting the inhibitory interaction between Bcl-2 and Beclin1.Finally, Nix associates with mitochondrial membranes and directlyinteracts with LC3 (Atg8).

Although the molecular regulation of mitophagy has not yet beencompletely elucidated, the mTOR/AMPK pathway is proposed to be a majorcheckpoint. AMPK, in addition to stimulating mitochondrial removalthrough autophagy, enhances the activity of sirtuin-1 (SIRT1) and itsdownstream target PGC-1α, resulting in stimulation of mitochondrialbiogenesis. Hence, through the activity of AMPK, mitophagy andmitochondrial biogenesis are coordinately regulated, maintaining ahealthy and functional pool of mitochondria in the cell.

Lipophagy is a recently recognized alternative pathway of lipidmetabolism in which intracellular lipid droplet triglycerides andcholesterol are taken up by autophagosomes and delivered to lysosomesfor degradation by acidic hydrolases, thereby releasing free fattyacids. Lipophagy, therefore, functions to regulate intracellular lipidstores, cellular levels of free lipids, such as fatty acids, and energyhomeostasis.

Xenophagy is a recently recognized mechanism of defense against varioustypes of intracellular pathogens, including Mycobacterium tuberculosis,Salmonella typhimurium, Legionella pneumophila, Brucella species,Chlamydia species, Coxiella burnetti, Listeria monocytogenes, Shigellaflexneri, Rickettsia species, Mycobacterium marinum, Burkholderiaspecies, and Francisella tularensis.

Microautophagy involves lysosomes directly engulfing cytoplasm byinvagination, protrusion, or septation of the lysosomal limitingmembrane.

Chaperone-Mediated Autophagy

Chaperone-mediated autophagy (CMA) concerns only those proteins thathave a consensus peptide sequence that can be recognized by the bindingof a hsc70-containing chaperone/co-chaperone complex. The CMAsubstrate/chaperone complex then moves to the lysosomes, where the CMAreceptor lysosome-associated membrane protein type-2a (LAMP-2A)recognizes it. The protein is unfolded and translocated across thelysosome membrane assisted by the lysosomal hsc70 on the other side.Thus, CMA substrates are translocated across the lysosomal membrane on aone-by-one basis, whereas in macroautophagy and microautophagy thesubstrates are engulfed or sequestered in bulk. Moreover, CMA degradesonly certain proteins and not organelles.

Exemplary Therapeutic Indications for Increased Autophagy

Compounds, compositions, and methods of the invention can be used totreat and prevent any of the following therapeutic indications forincreased autophagy.

Autophagy Protects Organisms from Metabolic Stress

Nutrient deprivation, growth factor depletion, and hypoxia can inducemetabolic stress leading to the induction of autophagy and to thegeneration of free amino acids and fatty acids. These can be recycled ina cell-autonomous fashion and be used for 1) de novo synthesis ofproteins important in the stress response, and 2) fueling the TCA cycleto maintain ATP function. The importance of this process is demonstratedin the inability of mice and C. elegans with deficiencies in the ATGproteins important for autophagy to resist starvation. Thus, a criticalrole for autophagy is the mobilization of intracellular energy resourcesto meet cellular and organismal demand for metabolic substrates.

Induction of Autophagy for Treatment of the Heart

Cardiomyocyte function and survival rely critically on the presence ofbasal levels of cardiomyocyte autophagy. Autophagic recycling of damagedcellular components in nutrient-rich conditions constitutes a majormeans of protein and organelle quality control, ridding the cell ofdefective (e.g., misfolded or oxidized) proteins and dysfunctionalorganelles. This fact is highlighted by the observation that abrogationof autophagic pathways in adult heart by conditional inactivation ofeither the Atg5 or Atg7 genes triggers rapid-onset cardiac hypertrophy,left ventricular dilation, and diminished cardiac output.

Danon disease, a condition marked by severe and progressive myopathy,stems from defective fusion of autophagosomes with lysosomes. In earlycardiac development, Atg5 disruption provokes in utero defects andembryonic lethality. At the other end of the age spectrum, age-relateddeclines in the efficiency of autophagic clearance likely contribute toprogressive accumulation of defective proteins and organelles whichultimately lead to functional deterioration over time. Normal aging isassociated with loss of cardiac function mainly due to impairedrelaxation during diastole. Varying formulations of caloric restriction(CR) can prolong lifespan and improve LV diastolic function; theunderlying mechanisms are believed to be the induction of autophagy.Together, these facts highlight the vital housekeeping role forcardiomyocyte autophagy as a mechanism of protein and organellesurveillance and quality control.

Autophagy can Improve Skeletal Muscle Function in Setting of MuscularAtrophy

Skeletal muscle adapts its capacity to levels of load and utilization. Acentral aspect of this adaption is the regulation of fiber remodelingthrough degeneration or regeneration of muscle fibers.

In the absence of muscle activity, muscular atrophy occurs, resulting indecreased muscular capacity. This atrophy has been shown to occur due toincreased levels of oxidative stress in disused muscle. Attenuation ofthis oxidative stress could lead to decreased atrophy.

The autophagy process, and in particular mitophagy are important inclearing damaged mitochondria and reducing the effects of increasedoxidative stress on muscle functional capacity. Failure of the autophagyprocess has been shown to be an important contributing factor to muscledisuse atrophy, by failing to remove damaged mitochondria. This decreasein mitochondria turnover leads to an accumulation of dysfunctionalorgans and ensuing muscle damage.

Preserving Autophagy Function During Aging can Improve Sarcopenia

Skeletal muscle atrophy and impaired muscle strength represent animportant health issue and may occur as a consequence of immobilization,disuse, injury, starvation, and aging. In particular, advanced age isineluctably accompanied by the loss of muscle mass and strength. Thiscondition, known as sarcopenia of aging, has significant effects onindividual health and impacts the severity of frailty. Moreover, poormuscular strength is highly predictive of disability and mortality, andgeneral weakness often results in the loss of independent living,thereby affecting individual quality of life and imposing a high burdenon healthcare expenditure. Aside from aging, skeletal muscle can undergosignificant atrophy following disuse.

Sarcopenia is characterized by a gradual loss of muscle proteins. Thesize of stable post-mitotic tissues, such as skeletal and cardiacmuscles, is regulated by protein turnover, and skeletal muscle isinfluenced by a balance between protein synthesis and degradation andthe turnover of contractile proteins. A key factor influencing thedevelopment of sarcopenia is the imbalance between the rates of proteinsynthesis and degradation. Protein degradation in skeletal muscle cellsis essentially mediated by the activity of two highly conservedpathways: the autophagic lysosomal pathway and the ubiquitin-proteasomepathway.

Recent studies have shown that the impaired autophagy seen in ATG7 nullmuscles is characterized by muscle atrophy, weakness, and features ofmyofiber degeneration. Consequently, autophagy has been found to beessential for myofiber maintenance and for the clearance of damagedproteins and altered organelles.

Autophagy, which is activated when skeletal muscle is under nutritionalstress (such as metabolic stress), plays a role in the cataboliccondition and in the degradation of macromolecules and organelles.Catabolic pathways are accelerated during exercise to supply energy andsubstrates to the muscle for continuation of contractions. It has beenwell established that the rates of amino acid (relatively small) andglucose oxidation are increased during endurance exercise, and increasedenergy consumption is likely required to induce autophagy. It has beenshown that autophagy is required for myofiber maintenance and for theclearance of damaged proteins and altered organelles.

Mild exercise has been shown to improve muscle function and decrease thedecline in muscle function observed in sarcopenia. These positivebenefits are at least in part due to an exercise induced improvement inthe autophagy process. In aging mice, the autophagy proteins LC3-II,Beclin-1, ATG7, and MuRF-1 significantly decrease with age in muscle.However, mice undergoing a training regimen during the aging processshow a significantly attenuated decrease in these autophagy proteins. Inoverweight older women, mild exercise has been shown to increase thetranscript levels of the autophagy regulators LCB3, Atg7, and LAMP-2 andthus improve the autophagy process. Thus, preservation of autophagy mayplay an important role in skeletal myocyte homeostasis and optimalmitochondrial turnover in aged muscle.

An age-related attenuation of autophagy has been shown and results in adiminished efficiency of protein degradation and the clearance ofdamaged organelles. A decrease in proteolytic activity has beenconsidered responsible, at least in part, for the accumulation ofdamaged cellular components in almost all tissues of aging organisms.

Improving Autophagy as a Therapeutic Target for Muscle DegenerativeDiseases

Muscular dystrophies are a group of genetic, hereditary muscle diseasescharacterized by defects in muscle proteins. These defects result inprogressive skeletal muscle damage accompanied by myofiber necrosis andchronic local inflammation, leading to substitution of myofibers byconnective and adipose tissue. In Duchenne muscular dystrophy (DMD), themost severe form of these diseases, the continuous and progressiveskeletal muscle damage leads to complete paralysis and death ofpatients, usually by respiratory and/or cardiac failure.

The therapeutic protocols currently in use, based on corticosteroidadministration, provide some delay in the progression of the disease,but they are associated with severe side effects. Therapies thatsubstitute corticosteroids or at least may act as corticosteroid-sparingdrugs are thus being actively pursued, and biological mechanismsrelevant to skeletal muscle homoeostasis are explored, in order toidentify new targets.

Autophagy is emerging as an important process that limits muscle damage.Inhibition/alteration of autophagy contributes to myofiber degenerationleading to accumulation of abnormal organelles. Mutations thatinactivate Jumpy, a phosphatase that counteracts the activation of VPS34for autophagosome formation and reduces autophagy, are associated with acentronuclear myopathy. This observation suggests that unbalancedautophagy is pathogenic in muscle degeneration. Likewise,hyperactivation of Akt as a consequence of muscle-specific deletion ofthe mammalian target of rapamycin (mTOR) leads to inhibition ofautophagy and to a muscle phenotype resembling the one observed inmuscular dystrophy. The validity of autophagy modulation as atherapeutic strategy has been shown in a mouse model of Ulrich myopathycharacterized by defective autophagy and accumulation of dysfunctionalorganelles. Forced reactivation of autophagy in these animals yielded abeneficial therapeutic response.

In vivo and ex vivo analyses have shown that autophagy is defective inboth the human (DMD) and mouse (mdx) muscular dystrophy and that suchdefect contributes to the pathogenesis of the disease. Muscle biopsiesfrom DMD patients have been shown to have significantly lower levels ofLC3 II and significant accumulation of p62, a protein known to beincorporated into autophagosomes and efficiently degraded, with respectto tissues from control, non-affected individuals.

A low protein diet has been shown in mice to lead to a prolongedinduction of autophagy. In mice with DMD fed a low protein diet, aninduction of autophagy leads to an improvement and management in thedisease progression. Significant improvements in muscle function havebeen observed with an improvement of whole body tension, reduced musclefibrosis, decreased collagen disposition, reduced accumulation ofdamaged organelles and reduced apoptosis of muscle fibers.

This demonstrates that induction of autophagy is an importanthomoeostatic mechanism that is disrupted in dystrophic muscles andindicates that novel therapeutic approaches aimed at reactivatingautophagy can serve as a valuable strategy to reduce muscle damage inDMD.

Autophagy Protects the Liver from Oxidative Stress and Disease

During liver diseases such as cancer and cirrhosis, the liver canundergo tissue hypoxia. This process has been shown to induce anautophagy process, which if inhibited resulted in increased apoptosis ofliver cells.

In α1-antitrypsin deficiency, the most common genetic cause of humanliver disease, there is significant chronic inflammation and eventualcarcinogenesis. In this disease, a point mutation occurs inα1-antitrypsin Z (ATZ) leading to improperly folding and accumulation ofaggregates. Deletion of ATG5 in hepatic cell lines lead to anaccumulation of the mutant ATZ protein, demonstrating the important rolefor autophagy in reducing the impact of liver disease.

Autophagy is Important in Limiting Ischemic Reperfusion Injury

With advancing age, patients are more likely to acquire primary andsecondary hepatic malignancies that are amenable to surgical resectionand transplantation. Though the elderly patients may be treatedsurgically, the aged liver has significantly decreased reparativecapacity following ischemia and reperfusion injury associated with theseoperations.

Ischemic preconditioning is the only promising strategy for improvingthe outcome of liver surgery, but its beneficial effects are limited toyoung patients. To date, no therapeutic strategy can suppress theage-dependent ischemia and reperfusion injury.

A reduction in autophagy has been observed in the old cells subjected toa severe stress such as ischemia followed by reperfusion. Studies haveshown that by overexpression of autophagy genes in aged livers of mice,autophagy was increased and hepatocyte cell survival was increased afterischemia and reperfusion. Consequently, defective autophagy has beenshown to be a causal mechanism for the age-dependent hepatic reperfusioninjury and that enhancement of autophagy has been demonstrated to offertherapeutic benefit and reducing age-mediated liver ischemia reperfusioninjury.

Autophagy in Intestinal Epithelial Cells as a Therapeutic Target

The intestinal epithelium interfaces directly with a diverse communityof bacteria that includes benign commensals, opportunistic pathogens,and overt pathogens, and consequently is the first line of defenseagainst bacterial invasion of host tissues. One means that theepithelial cells employ to defend themselves includes secretingantimicrobial proteins. Unfortunately, there are some intestinalpathogens, including Salmonella tyhpimurium or opportunisticallyinvasive commensal bacteria, such as Enterococcus faecalis, which canavoid this first line of defense and enter the epithelial cells.

Autophagy has been shown to be essential for the recognition anddegradation of intracellular pathogens, acting as an innate barrier toinfection. In cell culture, autophagy has been shown to limit thereplication of certain bacterial species.

It has been shown via genetic studies of inflammatory bowel disease(IBD) that autophagy plays an important role in the intestinal immunehomeostasis. IBD is a chronic inflammatory disease of the intestine thatarises from dysregulated interactions with resident microbiota.

Recently, it has been shown that polymorphisms in genes in theautophagic pathway are linked to Crohn's disease (CD). Crohn's diseaseis a chronic form of IBD that can affect any part of thegastrointestinal system, but is usually found in the colon or terminalileum. The average onset is at 27 years of age in humans, and is usuallypresent throughout the normal lifespan of the individual. It ischaracterized by severe colitis, strictures, and perianal fistulas,typically requiring surgery.

The chronic inflammatory process characteristic of CD requires theintensive interaction between intestinal epithelial cells and immunecompetent cells. In CD, there is an exaggerated immune response to theintestinal microbiota, characterized by an abnormal increase in Th17cells, which play a major role in autoimmunity, and a down-regulation ofTreg cells important for controlling the immune response.

It has recently been shown that intestinal epithelial cell autophagy isessential for mammalian intestinal defense against invasive bacteria.Autophagy in the epithelial cells protects against the dissemination ofinvasive bacteria. Following oral infection with the invasive pathogenSalmonella typhimurium as well as Enterococcus faecalis, mouseepithelial cells activate autophagy as a consequence of exposure tothese pathogens. Autophagy was also shown to be critical to limit theextra-intestinal spread of S. typhimurium. This indicates that autophagyis a key epithelial cell-autonomous mechanism of antibacterial defensethat protects against dissemination of intestinal bacteria.

The present invention provides the know-how to use compounds thatinclude urolithins and their precursors as enhancers of autophagy forthe administration to and the treatment of individuals with inflammatorybowel disease (IBD) or Crohn's disease (CD) and in need of increasingthe levels of autophagy in their in the epithelial cells of theintestine in order to treat either IBD or CD.

Autophagy is Important in Aging Cardiac Muscle

The effects of autophagy induction on improved outcome for ischemicinjury and muscle maintenance makes it especially relevant for cardiacmuscle maintenance and protection from injury. Cardiac muscle undergoesprogressive decline in mitochondrial function, similar to that observedin skeletal muscle, resulting in an increase in reactive oxygen species,as well as an increase in the accumulation of defective organelles. Theclearance of these damaged organelles by autophagy is important for themaintenance of cardiac muscle function. As autophagy decreases with age,promoting autophagy can serve to protect cardiac muscle function.

Cardiac muscle is also strongly exposed to ischemic episodes duringcardiac infarcts. The level of cardiac muscle damage that these ischemicepisodes produce is strongly dependent on the ability of the cells tomount an effective autophagy response to clear damaged organelles. Inaged animals, a defective autophagy response leads to an increase incardiac muscle damage after ischemic events. Thus, promotion ofautophagy during these acute events could serve to protect cardiacmuscle from damage.

Autophagy is Important in the Inflammatory Process

Due to the role of autophagy in clearing defective organelles, a defectin this process leads to a buildup of cellular debris and the inductionof apoptosis. Autophagy also plays an important role in defending theorganism against microbial pathogens by inducing their degradation.Additionally, autophagy plays an important role in the traffickingevents that activate innate and adaptive immunity.

The autophagic removal of apoptotic corpses is critical for preventingdanger signals that could lead to an inflammation response. In animpaired autophagy response, where apoptotic clearance is not efficientthe resulting induction of inflammation, could overcome tolerance toself-antigens leading to autoimmune diseases such as systemic lupuserythematosus. Thus, induction of autophagy could serve to decreaseinflammatory responses and the development of autoimmune diseases.

Applications of Autophagy for Treatment of Disorders of the Liver

A number of features of hepatocytes and the liver as a whole make thisorgan particularly dependent on autophagy. The liver is rather unique inits regenerative properties as while hepatocytes are normally in aquiescent state, they retain the ability to quickly enter the cell cyclewhen there is a loss of liver tissue due to injury or surgical removal.The lack of cell turnover makes hepatocytes particularly vulnerable tothe effects of impaired autophagy, as cells having long lives accumulatehigh levels of damaged organelles, protein aggregates, etc. that arenormally cleared by autophagy. This leads to cellular injury andpotentially to transformation.

Hepatocellular Lipid Metabolism

The liver serves as the second largest repository of stored lipids inthe body after adipose tissue. Hepatocytes are a major cellularstorehouse for neutral lipids in the form of triglycerides (TGs) andcholesterol esters contained in specialized organelles termed lipiddroplets (LD). Autophagy mediates the breakdown of intracellular LDstores through the process of lipophagy. This enables the hepatocytes torapidly mobilize their lipid stores in times of metabolic need. The lossof hepatocyte autophagy leads to a marked increase in hepatic TG andcholesterol content, indicating that lipophagy limits lipid accumulationby the liver in vivo. Also, lipophagy controls cellular energyhomeostasis by providing free fatty acids (FFA) from the breakdown ofTGs, which subsequently drives mitochondrial β-oxidation and cellularATP generation. It has been shown that the autophagosomal protein LC3,critical for autophagosome membrane formation, associates with LDs.

Autophagy Protects Against Hepatic Diseases

SERPINA1/α1-anti-trypsin deficiency (ATD) is the most common geneticcause of human liver disease in children. This disease is caused byhomozygosity for the SERPINA1/α1-antitrypsin Z allele SERPINA1-Z, apoint mutation, which renders the hepatic secretory glycoproteinSERPINA1 prone to misfolding, polymerization, and aggregation. Themutant SERPINA1-Z protein accumulates in hepatocytes and the levels ofSERPINA1 found in the blood and body fluids are reduced to 10-15% ofthose normally observed. Accumulation of mutant SERPINA1-Z in theendoplasmic reticulum (ER) of hepatocytes leads to liver damage by again-of-function. It has been shown that intracellular degradation ofSERPINA1-Z aggregates and polymers involves the autophagic pathway.

The drug carbamazepine, known to induce autophagy, was recently shown tobe effective in cell based and mouse model of ATD. Carbamazepineincreases autophagic degradation of SERPINA1-Z in cultured cells andwhen provided orally to the PiZ mouse model of ATD, it reduced thehepatic load of SERPINA1-Z. Additionally, inducing autophagy reducedhepatic fibrosis. Consequently, drugs enhancing autophagy are attractivecandidates for improving the liver disease that develops in somepatients with ATD.

The present invention provides the know-how to use compounds thatinclude urolithins and their precursors as enhancers of autophagy forthe treatment of individuals with ATD and in need of increasing thelevels of autophagy in their liver and hepatocytes in order to reduceliver toxicity.

Autophagy Protects Against Nonalcoholic Fatty Liver Disease

Nonalcoholic fatty liver disease (NAFLD) is an important component ofthe metabolic syndrome together with obesity and diabetes. NAFLDencompasses a spectrum of hepatic abnormalities that range from simplefatty liver or steatosis, to fatty liver with hepatocellular injury andinflammation, which is known as nonalcoholic steatohepatitis (NASH).NAFLD is now the most prevalent liver disease in the USA and accountsfor about 75% of all chronic liver diseases.

The most important role of autophagy in fatty liver disease could be toregulate the process of excessive lipid accumulation. In fact, mice witha hepatocyte-specific knockout of Atg7, a protein required forautophagy, consuming a high-fat diet led to a marked increase in liverTGs and cholesterol content, showing that autophagy defects can inducehepatic steatosis. When considering NASH, while its exact causes areunknown, free fatty acid (FFA)-induced lipotoxicity has been implicatedin the mechanisms of hepatocellular injury of this disease. Evidencepoints to the fact that hepatocyte autophagy renders the cells moreresistant to injury from FFA.

Autophagy is an attractive therapeutic target for the treatment andprevention of both NAFLD and NASH. Therapeutic intervention to increaseautophagy may reverse not only the hepatic manifestations of NAFLD,including hepatocellular steatosis and injury, but also some of theunderlying metabolic abnormalities of the disease via its effects oninsulin resistance. Additionally, treatment by increasing autophagy mayprevent common end-stage complications of NAFLD, such as hepatocellularcarcinoma.

The present invention provides the know-how to use compounds thatinclude urolithins and their precursors as enhancers of autophagy forthe treatment of individuals with NAFLD and in need of increasing thelevels of autophagy in their liver and hepatocytes in order to treatthese conditions.

Autophagy Protects Against Alcoholic Liver Disease

Alcoholic liver disease (ALD) is a major cause of chronic liver disease,and like NAFLD, has a wide spectrum of pathogenic features, that rangefrom steatosis to sever acute alcoholic hepatitis, fibrosis, cirrhosis,and hepatocellular carcinoma.

Autophagy has been shown to play a role in treating ALD. For example,induction of autophagy by administration of rapamycin significantlysuppresses acute alcohol-induced steatosis. Also, a common feature ofchronic alcohol abuse is the formation of hepatic protein aggregatesknown as Mallory-Denk bodies, which are cytosolic inclusion bodiesenriched with Krt8/keratin 8 and Krt18 and proteins that includeubiquitin. Rapamycin treatment significantly reduces the number ofMallory-Denk bodies in proteasome inhibitor-treated KRT8 transgenicmice.

Consequently, enhancing hepatic autophagy is an attractive target forimproving alcohol-induced liver disease. The present invention providesthe know-how to use compounds that include urolithins and theirprecursors as enhancers of autophagy for the treatment of individualswith ALD and in need of increasing the levels of autophagy in theirliver and hepatocytes in order to treat this condition.

Autophagy Protects Against Drug-Induced Liver Injury

Most drugs are metabolized and detoxified in the liver, making the liverthe principal target for drug damage. Liver injury due to drugs is acommon cause for the withdrawal of approved drugs on the market, and itis thought that drug-induced hepatotoxicity is responsible for more thanhalf of acute cases of liver failure. Acetaminophen, also known asparacetamol and N-acetyl-p-aminophenol (APAP), is a widely usedantipyretic and analgesic drug and is also the most common source ofsevere drug-induced hepatotoxicity. At therapeutic levels APAP is safe,but overdosing leads to toxicity mainly due to its reactive metabolite,N-acetyl-p-benzoquinone imine (NAPQI). NAPQI can deplete hepatic storesof glutathione (GSH), an intracellular antioxidant. Following thedepletion of GSH, NAPQI is known to react with cellular proteins as wellas mitochondrial proteins to form protein adducts. These APAP-inducedmitochondrial protein adducts can then lead to mitochondrial damage andsubsequent necrosis.

When autophagy is enhanced with rapamycin, APAP-induced necrosis issignificantly inhibited, both in cultured primary hepatocytes and in thelivers of mice. Treatment with rapamycin two hours after APAPadministration has been seen to significantly improve APAP-induced liverinjury, even though APAP metabolism and hepatic GSH depletion havealready occurred. This is particularly important as patients at risk forhepatotoxicity from an acute APAP overdose do not receive medical careuntil they are past the metabolic phase. Consequently, pharmacologicintervention targeting an enhancement of autophagy holds a potentialtherapeutic benefit for individuals with a risk of APAP hepatotoxicityfollowing an overdose.

The present invention provides the know how-to use compounds thatinclude urolithins and their precursors as enhancers of autophagy forthe treatment of individuals at risk of hepatotoxicity due to drug sideeffects and in need of increasing the levels of autophagy in their liverand hepatocytes in order to treat or prevent the potential drugtoxicity.

Autophagy is Important in Limiting Ischemia/Reperfusion Injury

Ischemia/reperfusion (I/R) injury is a causal factor contributing tomorbidity and mortality. The vulnerability of the liver to I/R injury isa major obstacle to liver resection and transplantation surgery wherereperfusion after sustained ischemia is unavoidable during hepatectomyand vascular reconstruction. Mitochondrial dysfunction is known to beone of the critical downstream events that lead to I/R-mediated celldeath.

Autophagy clears abnormal or dysfunctional mitochondria to ensure anoptimal cellular function and survival. With impaired or insufficientmitophagy, cells accumulate damaged mitochondria, which subsequentlyleads to uncontrolled ROS formation, mitochondrial DNA mutation,energetic failure, and ultimately cell death. Consequently, the failureof mitophagy to remove a small number of damaged mitochondria during I/Rcan have a significant impact on hepatocellular function and viability.Mitophagy is essential for hepatic function and survival following I/Rinjury.

While minimizing I/R injury plays an important role in the outcome oftransplanted young livers, aged livers are even more susceptible tonegative impact of I/R injury. In the case of aged livers, hepatocytesfail to respond to the I/R stress and upregulate their endogenousprotective autophagy response. Similar to young livers followingprolonged ischemia, aged livers after short-term ischemia accumulatedysfunctional mitochondria, undergo mitochondrial permeabilitytransition, and lose their viability soon after reperfusion.

Methods of enhancing autophagy, including pre-ischemia nutrientdepletion and over expression of pro-autophagy genes ATG7 or BECN1, leadto the suppression of the mitochondrial permeability transition andincreases hepatocyte survival following reperfusion.

This indicates that treatments with agents that induce autophagy in theliver will offer protection during a situation of I/R and help tominimize cellular injury. Such treatments are applicable in situationsof the transplantation of both young and aged livers. Treatments mayinvolve: (i) pre-treatments of the liver tissue ex vivo by perfusion ofthe liver with a solution that contains an inducer of autophagy; (ii)treatment of the liver donor with an autophagy inducer; or (iii)treatment of the liver recipient prior to, during the operation and/orimmediately after the surgical intervention. Of course, these treatmentmodalities may be applied individually or in any combination (forexample: 1 and 2; 2 and 3; 1 and 3; 1, 2, and 3).

The present invention provides the know-how to use compounds thatinclude urolithins and their precursors as enhancers of autophagy forthe treatment of individuals and their livers, that may be at risk ofI/R injury. These compounds may be provided orally or parenterally tothe donor or recipient, or provided in a preconditioning solution thatmay be applied to the resected liver tissue.

Autophagy and Osteoarthritis

Osteoarthritis (OA) is the most common aging-related joint pathology andis characterized by degradation of cartilage extracellular matrix (ECM)and reduced cartilage cellularity. Changes in the articular cartilageappear to be critical in OA initiation and progression. Chondrocytes arethe only cell population of adult articular cartilage. The capacity ofthe adult articular chondrocytes to regenerate the normal cartilagematrix architecture is limited and declines with aging, due to celldeath and abnormal responsiveness to anabolic stimuli. Articularcartilage is characterized by a very low rate of cell turnover and ithas been shown that autophagy play an important role in chondrocytecellular function and survival. In fact, autophagy is a constitutivelyactive and protective process for the maintenance of cartilagehomeostasis. Studies have shown both in joint aging and OA in humans andin mice that there is a reduction in the expression of autophagyregulators, which was accompanied by an increase in chondrocyteapoptosis. Compromised autophagy is thought to contribute to thedevelopment of OA. It has been shown that treatment with the compoundrapamycin, a known inducer of autophagy, has been able to increaseactivation of LC3 in cartilage in an animal model of OA and consequentlyreduce the severity of articular cartilage degradation. In the presentinvention, urolithins and their precursors have been shown to increasethe levels of autophagy in tissues following oral consumption, makingthem ideal candidates for the treatment and reduction of the severity ofosteoarthritis in young and aging humans and mammals.

Metabolic Syndrome, Diabetes, and Obesity

Compounds and methods of the invention are useful in the treatment andprevention of metabolic syndrome, type 2 diabetes mellitus, and obesity.As used herein, the term “metabolic syndrome” refers to a combination ofmedical disorders that, when occurring together, increase the risk ofdeveloping cardiovascular disease and diabetes. It affects one in fivepeople in the United States and prevalence increases with age. Somestudies have shown the prevalence in the United States to be anestimated 25% of the population. In accordance with the InternationalDiabetes Foundation consensus worldwide definition (2006), metabolicsyndrome is central obesity plus any two of the following:

-   -   Raised triglycerides: >150 mg/dL (1.7 mmol/L), or specific        treatment for this lipid abnormality;    -   Reduced HDL cholesterol: <40 mg/dL (1.03 mmol/L) in males, <50        mg/dL (1.29 mmol/L) in females, or specific treatment for this        lipid abnormality;    -   Raised blood pressure: systolic BP>130 or diastolic BP>85 mm Hg,        or treatment of previously diagnosed hypertension; and    -   Raised fasting plasma glucose: (FPG)>100 mg/dL (5.6 mmol/L), or        previously diagnosed type 2 diabetes.

Autophagy and Neurodegenerative Diseases

In neurodegenerative diseases, brain tissue accumulates autophagosomes,demonstrating an increase in autophagy, which in model organisms hasbeen shown to have a protective effect. It plays an important role inclearing the misfolded proteins that accumulate as a result of severalneurodegenerative diseases. These include proteins that have polyQrepeats as seen as in Huntington's diseases and spinocerebellar ataxia,mutant α-synucleins involved in Parkinson's as well as tau aggregates.

Knockdown of ATG genes important in autophagy in C. elegans resulted inincreased aggregate formation and toxicity of PolyQ proteins. InAlzheimer's disease the autophagy process is impaired as a result of adefect in autophagosomal maturation that could be an important reasonfor aggregate accumulation. By contrast, autophagy induction byrapamycin in both Drosophila and mouse models of polyQ disease protectedthese animals from neurotoxicity. These results demonstrate thatautophagy induction can have a protective role in neuronal cells againstneurodegeneration.

The development of neurodegenerative diseases in patients implies thatautophagy can reach a saturation point in which the ability to degrademutant protein aggregates is exceeded. Thus, promotion of autophagycould help in delaying the onset of neurodegeneration disease.

Cognitive Disorder

Compounds and methods of the invention are useful for treating acognitive disorder. As used herein, a cognitive disorder refers to anycondition that impairs cognitive function. In one embodiment, “cognitivedisorder” refers to any one or more of delirium, dementia, learningdisorder, attention deficit disorder (ADD), and attention deficithyperactivity disorder (ADHD). In one embodiment, the cognitive disorderis a learning disorder. In one embodiment, the cognitive disorder isattention deficit disorder (ADD). In one embodiment, the cognitivedisorder is attention deficit hyperactivity disorder (ADHD).

Compounds and methods of the invention are useful for improvingcognitive function, even in the absence of a cognitive disorder. As usedherein, “cognitive function” refers to any mental process that involvessymbolic operations, e.g., perception, memory, attention, speechcomprehension, speech generation, reading comprehension, creation ofimagery, learning, and reasoning. In one embodiment, “cognitivefunction” refers to any one or more of perception, memory, attention,and reasoning. In one embodiment, “cognitive function” refers to memory.

Methods for measuring cognitive function are well known and can include,for example, individual or battery tests for any aspect of cognitivefunction. One such test is the Prudhoe Cognitive Function Test.Margallo-Lana et al. (2003) J Intellect Disability Res. 47:488-492.Another such test is the Mini Mental State Exam (MMSE), which isdesigned to assess orientation to time and place, registration,attention and calculation, recall, language use and comprehension,repetition, and complex commands. Folstein et al. (1975) J Psych Res.12:189-198. Other tests useful for measuring cognitive function includethe Alzheimer Disease Assessment Scale-Cognitive (ADAS-Cog) (Rosen etal. (1984) Am J Psychiatry. 141(11):1356-64) and the CambridgeNeuropsychological Test Automated Battery (CANTAB) (Robbins et al.(1994) Dementia. 5(5):266-81). Such tests can be used to assesscognitive function in an objective manner, so that changes in cognitivefunction, for example in response to treatment in accordance withmethods of the invention, can be measured and compared.

Protein Misfolding and Aggregation

These diseases and disorders, which are collectively referred to hereinas “amyloid-related diseases”, fall into two main categories: (a) thosewhich affect the brain and other parts of the central nervous system;and (b) those which affect other organs or tissues around the body.

Examples of amyloid-related diseases which fall under these twocategories are listed in the following two sections; however, many otherexamples of rare, hereditary amyloid-related diseases are known whichare not included here, and additional forms of amyloid-related diseaseare likely to be discovered in future.

Neurodegenerative Diseases Associated with Amyloidosis

Many different neurodegenerative diseases are associated with themisfolding and aggregation of a specific protein or peptide in aparticular part of the brain, or elsewhere in the central nervoussystem, depending on the specific disease. Examples of such diseasesfollow.

Various forms of Alzheimer's disease (AD) as well as Down's syndrome,hereditary cerebral hemorrhage with amyloidosis (HCHWA, Dutch type),cerebral amyloid angiopathy, and possibly also mild cognitive impairmentand other forms of dementia are associated with the aggregation of a40/42-residue peptide called β-amyloid, Aβ(1-40) or Aβ(1-42), whichforms insoluble amyloid fibers and plaques in the cerebral cortex,hippocampus or elsewhere in the brain, depending on the specificdisease. Alzheimer's disease is also associated with the formation ofneurofibrillary tangles by aggregation of a hyperphosphorylated proteincalled tau, which also occurs in frontotemporal dementia (Pick'sdisease).

Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiplesystem atrophy (MSA) are associated with the aggregation of a proteincalled α-synuclein, which results in the formation of insolubleinclusions called Lewy bodies. Huntington's disease (HD), spinal andbulbar muscular atrophy (SBMA, also known as Kennedy's disease),dentatorubral pallidoluysian atrophy (DRPLA), different forms ofspinocerebellar ataxia (SCA, types 1, 2, 3, 6 and 7), and possiblyseveral other inheritable neurodegenerative diseases are associated withthe aggregation of various proteins and peptides that contain abnormallyexpanded glutamine repeats (extended tracts of polyglutamine).Creutzfeldt-Jakob disease (CJD), bovine spongiform encephalopathy (BSE)in cows, scrapie in sheep, kuru, Gerstmann-Straussler-Scheinker disease(GSS), fatal familial insomnia, and possibly all other forms oftransmissible encephalopathy are associated with the self-propagatingmisfolding and aggregation of prion proteins.

Amyotrophic lateral sclerosis (ALS), and possibly also some other formsof motor neuron disease (MND) are associated with the aggregation of aprotein called superoxide dismutase.

Familial British dementia (FBD) and familial Danish dementia (FDD),respectively, are associated with aggregation of the ABri and ADanpeptide sequences derived from the BRI protein.

Hereditary cerebral hemorrhage with amyloidosis (HCHWA, Icelandic type)is associated with the aggregation of a protein called cystatin C.

Systemic Diseases Associated with Amyloidosis

In addition to the neurodegenerative diseases listed above, a widevariety of systemic ageing-related or degenerative diseases areassociated with the misfolding and aggregation of a particular proteinor peptide in various other tissues around the body (i.e., outside ofthe brain). Examples of such diseases follow.

Type II diabetes mellitus (also known as adult-onset diabetes, ornon-insulin dependent diabetes mellitus) is associated with theaggregation of a 37-residue peptide called the islet amyloid polypeptide(IAPP, or “amylin”), which forms insoluble deposits that are associatedwith the progressive destruction of insulin-producing β cells in theislets of Langerhans within the pancreas.

Dialysis-related amyloidosis (DRA) and prostatic amyloid are associatedwith the aggregation of a protein called β₂-microglobulin, either inbones, joints and tendons in DRA, which develops during prolongedperiods of hemodialysis, or within the prostate in the case of prostaticamyloid.

Primary systemic amyloidosis, systemic AL amyloidosis andmyeloma-associated amyloidosis are associated with the aggregation ofimmunoglobulin light chain (or in some cases immunoglobulin heavy chain)into insoluble amyloid deposits, which gradually accumulate in variousmajor organs such as the liver, kidneys, heart and gastrointestinal (GI)tract.

Reactive systemic AA amyloidosis, secondary systemic amyloidosis,familial Mediterranean fever, and chronic inflammatory disease areassociated with the aggregation of serum amyloid A protein, which formsinsoluble amyloid deposits that accumulate in major organs such as theliver, kidneys and splee. Senile systemic amyloidosis (SSA), familialamyloid polyneuropathy (FAP) and familial amyloid cardiomyopathy (FAC)are associated with the misfolding and aggregation of different mutantsof transthyretin protein (TTR), which form insoluble inclusions invarious organs and tissues such as the heart (especially in FAC),peripheral nerves (especially in FAP) and gastrointestinal (GI) tract.Another form of familial amyloid polyneuropathy (FAP, type II) isassociated with the aggregation of apolipoprotein AI in the peripheralnerves. Familial visceral amyloidosis and hereditary non-neuropathicsystemic amyloidosis are associated with misfolding and aggregation ofvarious mutants of lysozyme, which form insoluble deposits in majororgans such as the liver, kidneys and spleen.

Finnish hereditary systemic amyloidosis is associated with aggregationof a protein called gelsolin in the eyes (particularly in the cornea).

Fibrinogen α-chain amyloidosis is associated with aggregation of thefibrinogen A α-chain, which forms insoluble amyloid deposits in variousorgans, such as the liver and kidneys.

Insulin-related amyloidosis occurs by the aggregation of insulin at thesite of injection in diabetics.

Medullary carcinoma of the thyroid is associated with the aggregation ofcalcitonin in surrounding tissues.

Isolated atrial amyloidosis is associated with the aggregation of atrialnatriuretic peptide (ANP) in the heart.

Various forms of cataract are associated with the aggregation ofγ-crystallin proteins in the lens of the eyes.

Autophagy and Endothelial Cell Function and Associated Diseases

Endothelial Cell Dysfunction

Global endothelial cell dysfunction occurs in several diverse diseasessuch as diabetes, hypertension, chronic kidney disease, andatherosclerosis. In these diseases endothelial cell dysfunction isthought to occur as a result of stress-induced premature senencense(SIPS). SIPS is characterized by subverted autophagy and lysosomaldysfunction, with the accumulation of autolysosomal vacuoles.

Endothelial cell dysfunction also occurs as a result of aging with anincreased incidence of cardiovascular diseases. This increase in celldysfunction correlates with a decrease in autophagy. In older humans,expression of autophagy markers in arterial endothelial cells wasimpaired by 50% (P<0.05) and was associated with a 30% (P<0.05)reduction in arterial endothelium-dependent dilatation (EDD). Similarly,in C57BL/6 control mice aging was associated with a 40% decrease(P<0.05) in arterial markers of autophagy and a 25% reduction (P<0.05)in EDD, demonstrating that impaired autophagy is a cause of age-relatedarterial dysfunction.

In old mice, treatment with the autophagy-enhancing agent trehaloserestored expression of autophagy markers, rescued NO-mediated EDD byreducing oxidative stress, and normalized inflammatory cytokineexpression. The present invention provides the know-how to use compoundsthat include urolithins and their precursors as enhancers of autophagyfor the treatment of individuals having health conditions linked toendothelial cell dysfunction and in need thereof.

Endothelial Cell Injury

Endothelial cell injury can occur as a result of disease processes suchas sickle cell anemia or thalassemia in which pathologically high levelsof heme and iron release can occur. Severe skeletal muscle damage, aswell as cardiac ischemia injury results in the release of the hemeprotein, myoglobin, which also results in endothelial cell injury. Thisdamage to the vascular endothelial cells can lead to vasculardysfunction and an increase in cardiovascular complications. Endothelialcell injury caused by heme toxicity is associated with a progressivedecrease in endothelial cell mitochondrial membrane potential, leadingto apoptosis.

Micro- and macro-vascular complications are commonly seen in diabeticpatients, and endothelial dysfunction contributes to the development andprogression of the complications. Abnormal functions in endothelialcells lead to the increase in vascular tension and atherosclerosis,followed by systemic hypertension as well as increased incidence ofischemia and stroke in diabetic patients. Mitochondrial dysfunctionappears to be central to the vascular endothelial dysfunction. Enhancedmitochondrial fission and/or attenuated fusion leads to mitochondrialfragmentation and disruption of the endothelial physiological function.Abnormal mitochondrial biogenesis and disturbance of mitochondrialautophagy increase the accumulation of damaged mitochondria, such asirreversibly depolarized or leaky mitochondria, and facilitate celldeath. Augmented mitochondrial ROS production and Ca²⁺ overload inmitochondria not only cause the maladaptive effect on the endothelialfunction, but also are potentially detrimental to cell survival.

Endothelial cell injury can also result from cardiac procedures such asangioplasty, bypass surgery, and valve replacement. Upregulation ofautophagy should lead to a reduction in the injury to the associatedendothelial cells.

Strategies that increase autophagy would have clear therapeuticpotential. The present invention provides the know-how to use compoundsthat include urolithins and their precursors as enhancers of autophagyfor the treatment of individuals having health conditions linked toendothelial cell injury resulting from disease processes such asdiabetes, sickle cell anemia, or thalassemia, as well as protectingendothelial cells from the more acute effects of severe muscle injury.

Autophagy and Cancer

Autophagy and cancer have similar regulatory pathways, with severaltumor suppression genes such as PTEN, TSC1 and TSC2 leading to theupstream inhibition of TOR signaling, leading to the stimulation ofautophagy. Additionally, the autophagy protein, Beclin 1 has beenidentified as a tumor suppressor deleted in many human cancers. Theseresults demonstrate that autophagy plays an important role in tumorsuppression.

Aging

By far the greatest risk factor for neurodegenerative diseases, such asAlzheimer's disease (AD), Parkinson's disease (PD), and amyotrophiclateral sclerosis (ALS), is aging. Mitochondria have been thought tocontribute to aging through the accumulation of mitochondrial DNA(mtDNA) mutations and net production of reactive oxygen species (ROS).Although most mitochondrial proteins are encoded by the nuclear genome,mitochondria contain many copies of their own DNA. Human mtDNA is acircular molecule of 16,569 base pairs that encodes 13 polypeptidecomponents of the respiratory chain, as well as the rRNAs and tRNAsnecessary to support intramitochondrial protein synthesis using its owngenetic code. Inherited mutations in mtDNA are known to cause a varietyof diseases, most of which affect the brain and muscles—tissues withhigh energy requirements. It has been hypothesized that somatic mtDNAmutations acquired during aging contribute to the physiological declinethat occurs with aging and aging-related neurodegeneration. It is wellestablished that mtDNA accumulates mutations with aging, especiallylarge-scale deletions and point mutations. In the mtDNA control region,point mutations at specific sites can accumulate to high levels incertain tissues: T414G in cultured fibroblasts,

A189G and T408A in muscle, and C150T in white blood cells. However,these control-region “hot spots” have not been observed in the brain.Point mutations at individual nucleotides seem to occur at low levels inthe brain, although the overall level may be high. Using a polymerasechain reaction (PCR)-cloning-sequencing strategy, it was found that theaverage level of point mutations in two protein-coding regions of brainmtDNA from elderly subjects was ˜2 mutations per 10 kb. Noncodingregions, which may be under less selection pressure, potentiallyaccumulate between twice and four times as many. The accumulation ofthese deletions and point mutations with aging correlates with declinein mitochondrial function. For example, a negative correlation has beenfound between brain cytochrome oxidase activity and increasedpoint-mutation levels in a cytochrome oxidase gene (GM).

Net production of ROS is another important mechanism by whichmitochondria are thought to contribute to aging. Mitochondria containmultiple electron carriers capable of producing ROS, as well as anextensive network of antioxidant defenses. Mitochondrial insults,including oxidative damage itself, can cause an imbalance between ROSproduction and removal, resulting in net ROS production. The importanceto aging of net mitochondrial ROS production is supported byobservations that enhancing mitochondrial antioxidant defenses canincrease longevity. In Drosophila, overexpression of the mitochondrialantioxidant enzymes manganese superoxide dismutase (MnSOD) andmethionine sulfoxide reductase prolongs lifespan. This strategy is mostsuccessful in short-lived strains of Drosophila, and has no effect inalready long-lived strains. However, it has recently been shown thatoverexpression of catalase experimentally targeted to mitochondriaincreased lifespan in an already long-lived mouse strain.

Improving Activity During Aging

Activity in animals is driven largely by circadian rhythm and issynchronized to the environment. Disruption of the circadian rhythm or adesynchronization with the environment can lead to increase in nighttimewakefulness or daytime naps.

Normal aging is accompanied by declining locomotor activity, alteredcircadian rhythms, as well as altered sleep and food intake patterns.These effects lead to a decrease in alertness and vigilance decreases inthe elderly, leading to an increase in nighttime wakefulness, as well asan increase in daytime naps. Activity patterns can also be disrupted ina similar way by disease, such as Alzheimer's Disease.

Age-dependent changes in activity rhythms are also observed in otheranimals, for example, rats, hamsters, mice and dogs, with an increase infragmentation and a decrease in synchronization with the environment.These age-dependent circadian disruptions have been linked to thedegeneration of the suprachiasmatic nucleus of the hypothalamus. Sleepdisruption in both humans and rodents have been shown to contribute toage-dependent cognitive dysfunction.

The increasing disruption of circadian rhythms is accompanied by agradual decline in motor activity with age in several species, includinghumans, mice, monkeys, and dogs. Of particular note, the daytimeactivity of senior dogs (>10 years of age) declines as compared to youngand middle-aged dogs. These changes in activity can be monitored bydevices intended to measure activity, for example, by means of anaccelerometer or by employing motion sensing cameras.

Many of the disruptions seen in aging as a result of decreased activityare also observed in younger populations where cultural trends haveresulted in decreased activity, accompanied with increased caloricintake, leading to an obesity epidemic. The resulting caloric imbalancehas led to an increase in several disease conditions such as type 2diabetes, colon cancer, and metabolic syndrome, as well as mental healthissues. Several prospective cohort studies and meta-analysis in humanshave shown that physical inactivity is associated with an elevated riskfor the development of metabolic syndrome, type 2 diabetes,hypertension, coronary artery disease, stroke, and cardiovasculardisease.

Both humans and animals, particularly dogs, would benefit from thepresent invention and its ability to improve activity both during theyouth and aging periods of life.

In one embodiment, the urolithin or precursor would increase activity ofthe recipient, human or animal. In yet another embodiment, the increasein activity is an increase by 1% to 100%. For example, the activity maybe increased by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%. In certainembodiments, the increase in activity is an increase of 5-10%, 10-15%,15-20%, 20-25%, 25-30%, 30-35%, 35-40%, 40-45%, 45-50%, 50-55%, 55-60%,60-65%, 65-70%, 70-75%, 75-80%, 80-85%, 85-90%, 90-95%, and 95-100%.

In one embodiment, treatment by urolithin or a precursor thereof wouldincrease activity and lead to a reduction in risk of metabolic syndrome.In one embodiment, treatment by urolithin or a precursor thereof wouldincrease activity and lead to a reduction in risk of type 2 diabetes. Inone embodiment, treatment by urolithin or a precursor thereof wouldincrease activity and lead to a reduction in risk of hypertension,coronary artery disease, stroke, and cardiovascular disease. In oneembodiment, treatment by urolithin or a precursor thereof would increaseactivity and improve cognitive function.

Mood Disorders

Compounds and methods of the invention are useful for treating a mooddisorder (also known as an affective disorder). As used herein, a “mooddisorder” refers to a disturbance in emotional state, such as is setforth in the Diagnostic and Statistical Manual of Mental Disorders,published by the American Psychiatric Association. Mood disordersinclude but are not limited to major depression, postpartum depression,dysthymia, and bipolar disorder. In one embodiment, the mood disorder ismajor depression.

Compounds and methods of the invention are useful for treating orpreventing a stress-induced or stress-related mood disorder. As usedherein, a “stress-induced or stress-related mood disorder” refers to adisturbance in emotional state that is induced or related to stress.Such mood disorders are sometimes referred to as reactive mood disordersand are to be distinguished from other mood disorders, e.g., so-calledorganic mood disorders.

Compounds and methods of the invention are useful for treating ananxiety disorder. As used herein, an “anxiety disorder” refers to adysfunctional state of fear and anxiety, e.g., fear and anxiety that isout of proportion to a stressful situation or the anticipation of astressful situation. In one embodiment, an anxiety disorder is any oneor combination of generalized anxiety disorder, panic disorder, panicdisorder with agoraphobia, agoraphobia, social anxiety disorder,obsessive-compulsive disorder, and post-traumatic stress disorder. Inone embodiment, an anxiety disorder is any one or combination ofgeneralized anxiety disorder, obsessive-compulsive disorder, panicdisorder, post-traumatic stress disorder, and social anxiety disorder.In one embodiment, an anxiety disorder is generalized stress disorder.In one embodiment, an anxiety disorder is post-traumatic stressdisorder. In one embodiment, an anxiety disorder is a stress-inducedanxiety disorder.

Compounds and methods of the invention are useful for treating orpreventing a stress-induced or stress-related anxiety disorder. As usedherein, a “stress-induced or stress-related anxiety disorder” refers toa dysfunctional state of fear and anxiety that is induced or related tostress. Such anxiety disorders are sometimes referred to as reactiveanxiety disorders and are to be distinguished from other anxietydisorders, e.g., so-called organic anxiety disorders.

For purposes of this invention, each of the foregoing diseases orconditions is associated with, or characterized by, reduced or decreasedautophagy, or would benefit from increased autophagy, and thereforewould benefit from the administration of urolithins and theirprecursors.

Table 1 summarizes some of the types of cells affected by autophagy andthe beneficial effects of autophagy in those cells.

TABLE 1 Health Benefits of Increased Autophagy Cell Type ConditionImproved by Autophagy All cells Maintenance of the amino acid poolduring starvation Anti-aging Tumor suppression Clearance ofintracellular microbes Clearance of protein aggregates NeuronsPrevention of neurodegeneration Alleviation of symptoms related toneurodegeneration Clearance of protein aggregates Acinar cells ofImproved outcome in acute pancreatitis the pancreas Smooth muscle cellsImproved outcome in various heart conditions such as heart disease,cardiac hypertrophy, left ventricular dilation, Danon diseaseAmelioration of cardiomyopathy resulting from diabetes Preventingcardiac deterioration with age Improvement outcome fromischemia/reperfusion injury Protection of cells from ischemia andhypoxic conditions resulting from myocardial function Improved cardiacoutput Improved left ventricular diastolic function Improvement in bloodpressure Protection of cells during cardiac procedures such asangioplasty, bypass, valve replacement Intestinal epithelial Improvementof immune response to cells intracellular bacteria; Crohn's disease;inflammatory bowel disease (IBD) Podocytes in kidney Podocyte resistanceto injury, which can be caused by the following diseases: minimal changedisease, focal segmental glomerulosclerosis, diabetic nephropathy,membrane glomerulopathy, lupus nephritis, and experimental glomerulardisease Skeletal muscle cells Protection against glucose intolerance,leptin resistance, high cholesterol and triglycerides in high fat dietsImproved muscle function Reduction in sarcopenia Improved balance andcoordination Improvement in muscle strength Increase in muscle massReduction in muscle atrophy Improvement in muscle endurance Improvedoutcome for muscular dystrophy Improvement in muscle recovery followingexercise Protection against muscle damage Liver tissue Clearance ofmutant alpha-antitrypsin Improved outcome in nonalcoholic fatty liverdisease (NAFLD), non-alcoholic steatohepatitis (NASH), and alcoholicliver disease (ALD) Liver hepatocyte Protection against glucoseintolerance, leptin resistance, high cholesterol and triglycerides inhigh fat diets Treatment of obesity and type II diabetes Improvementoutcome from ischemia/reperfusion injury Protection against drug inducedliver injury Protection of liver tissue Pancreatic beta cells Protectionagainst glucose intolerance, leptin resistance, high cholesterol andtriglycerides in high fat diets Adipocytes Protection against glucoseintolerance, leptin resistance, high cholesterol and triglycerides inhigh fat diets Treatment of obesity and type II diabetes Endothelialcells Improves endothelial cell dysfunction which can occur as a resultof diabetes, hypertension, chronic kidney disease, atherosclerosis andaging Protects against endothelial cell injury resulting from diseaseprocesses such as, diabetes, sickle cell anemia or thalassemia, as wellas from severe muscle injury. Protection of cells from ischemia andhypoxic conditions resulting from myocardial function Improvedangiogenesis Protection of cells during cardiac procedures such asangioplasty, bypass, valve replacement Chondrocytes/cartilage Reductionin osteoarthritis in joints Osteoblasts Improvement of tissue mineraldensity Increased bone strength Lens epithelial cells Reduction inage-related cataracts Retinal cells Reduction in age related maculardegeneration Ganglion cell layer Protection from diabetic retinopathyInner nuclear layer Outer nuclear layer Retinal pigment epithelial cellsRetinal Improved outcome from intraocular photoreceptors inflammation oruveitis Reduction in photoreceptor injury from uveitis Glaucomatoustissue Reduction in glaucomatous neurodegeneration KeratinocytesTreatment of intracellular skin infections, warts, psoriasis Protectionof skin from environmental insults i.e. UV light, anti-aging Reductionof skin injury from excessive lipid oxidation commonly observed in agedand diseased skin Lung cells Clearance of protein aggregates inpulmonary cells Improved outcome from pulmonary emphysema cause byal-antrypsin Chronic obstructive pulmonary disorder Alveolar Improvedoutcome for chronic obstructive macrophages pulmonary disease Airwayepithelial cells Reduction in protein aggregates in cystic fibrosisImproved clearance of aggresomes that accumulate mutant cystic fibrosistransmembrane conductance regulator (CFTR) protein Improved outcome forhuman idiopathic pulmonary fibrosis Immune cells Reduction ofauto-immune disorders Improved immune response to infection frompathogens

Exemplary Compounds of the Invention

Compounds of the invention include certain urolithins and precursorsthereof. These compounds can be used to practice any of the methodsherein, including but not limited to increasing autophagy or longevity,and for the treatment or prevention of the various diseases andconditions described herein.

In certain embodiments, the invention relates to urolithins. As usedherein, a “urolithin” refers to a compound of Formula I:

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

As used herein, the term “alkyl” refers to a straight chain or branched,noncyclic or cyclic, unsaturated or saturated aliphatic hydrocarbonradical containing carbon atoms. Representative saturated straight chainalkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, andthe like; while saturated branched alkyls include isopropyl, sec-butyl,isobutyl, tert-butyl, isopentyl, and the like. Representative saturatedcyclic alkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,and the like; while unsaturated cyclic alkyls include cyclopentenyl andcyclohexenyl, and the like. Unsaturated alkyls contain at least onedouble or triple bond between adjacent carbon atoms (referred to as an“alkenyl” or “alkynyl”, respectively). Representative straight chain andbranched alkenyls include ethylenyl, propylenyl, 1-butenyl, 2-butenyl,isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and the like; whilerepresentative straight chain and branched alkynyls include acetylenyl,propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl,3-methyl-1-butynyl, and the like.

As used herein, the term “aryl” refers to a hydrocarbon ring systemradical comprising hydrogen, 6 to 18 carbon atoms and at least onearomatic ring. For purposes of this invention, the aryl radical may be amonocyclic, bicyclic, tricyclic or tetracyclic ring system, which mayinclude fused or bridged ring systems. Aryl radicals include, but arenot limited to, aryl radicals derived from aceanthrylene,acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane,indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene, andtriphenylene. Unless stated otherwise specifically in the specification,the term “aryl” or the prefix “ar-” (such as in “aralkyl”) is meant toinclude aryl radicals that are optionally substituted.

As used herein, the term “monosaccharide” refers to a simple sugar ofthe formula (CH₂O)_(n). The monosaccharides can be straight-chain orring systems, and can include a saccharose unit of the formula—CH(OH)—C(═O)—. Examples of monosaccharides include erythrose, threose,ribose, arabinose, xylose, lyxose, allose, altrose, glucose, mannose,gulose, idose, galactose, talose, erythulose, ribulose, xyulose,psicose, fructose, sorbose, tagatose, erythropentulose, threopentulose,glycerotetrulose, glucopyranose, fructofuranose. In certain embodiments,monosaccharide refers to glucopyranose.

As used herein, the term “oligosaccharide” refers to saccharideconsisting of at least two, up to 10 glycosidically linkedmonosaccharide units, preferably of 2 to 8 monosaccharide units, morepreferably of 2 to 7 monosaccharide units, and even more preferably of 2to 6 monosaccharide units or of 2 to 5 monosaccharide units.

The term “substituted” as used herein (for example, in the context of asubstituted heterocyclyl or substituted aryl) means that at least onehydrogen atom is replaced with a substituent. “Substituents” within thecontext of this invention include halogen, hydroxy, oxo, cyano, nitro,imino, thioxo, amino, alkylamino, dialkylamino, alkyl, alkoxy,alkylthio, haloalkyl, aryl, aralkyl, heteroaryl, heteroarylalkyl,heterocycle and heterocyclealkyl, as well as —NR_(a)R_(b),—NR_(a)C(═O)R_(b), —NR_(a)C(═O)NR_(a)NR_(b),—NR_(a)C(═O)OR_(b)—NR_(a)SO₂R_(b), —C(═O)R_(a), —C(═O)OR_(a),—C(═O)NR_(a)R_(b), —OC(═O)NR_(a)R_(b), —OR_(a), —SR_(a), —SOR_(a),—S(═O)₂R_(a), —OS(═O)₂R_(a), —S(═O)₂OR_(a), ═NSO₂R_(a) and—SO₂NR_(a)R_(b). In the foregoing, R_(a) and R_(b) in this context maybe the same or different and independently hydrogen, alkyl, haloalkyl,cycloalkyl, aryl, aralkyl, heterocyclyl. In addition, the foregoingsubstituents may be further substituted with one or more of the abovesubstituents.

In one embodiment, the urolithin is urolithin A.

In one embodiment, the urolithin is urolithin B.

In one embodiment, the urolithin is urolithin C.

In one embodiment, the urolithin is urolithin D.

In one embodiment, a “urolithin” refers to any one or combination ofurolithin A, urolithin B, urolithin C, and urolithin D (see, forexample, FIG. 2 and FIG. 3 ). In one embodiment, a urolithin isurolithin A, urolithin B, urolithin C, urolithin D, or any combinationof urolithin A, urolithin B, urolithin C, and urolithin D. In oneembodiment, a urolithin is urolithin A, urolithin B, urolithin C, or anycombination of urolithin A, urolithin B, and urolithin C. In oneembodiment, a urolithin is urolithin A, urolithin B, or a combination ofurolithin A and urolithin B. In one embodiment, a urolithin is urolithinA.

In one embodiment, a urolithin is provided as an isolated urolithin,e.g., isolated from a natural source or prepared by total synthesis.Isolated urolithins may be synthesized de novo. See Examples 1-4.

In one embodiment, a urolithin is provided as a purified urolithin.

In one embodiment, a “urolithin” as used herein is or can include aglucuronated, methylated, or sulfated urolithin.

In certain embodiments, the invention relates to a compound of FormulaII

wherein

X¹, X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are independently selected from thegroup consisting of H and OH;

with the proviso that the compound is not a compound of Formula IIwherein

X¹, X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X¹ is OH, and X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X² is OH, and X¹, X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H (urolithin B);

X³ is OH, and X¹, X², X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X⁴ is OH, and X¹, X², X³, X⁵, X⁶, X⁷, and X⁸ are H;

X⁵ is OH, and X¹, X², X³, X⁴, X⁶, X⁷, and X⁸ are H;

X⁶ is OH, and X¹, X², X³, X⁴, X⁵, X⁷, and X⁸ are H;

X⁷ is OH, and X¹, X², X³, X⁴, X⁵, X⁶, and X⁸ are H;

X⁸ is OH, and X¹, X², X³, X⁴, X⁵, X⁶, and X⁷ are H;

X¹ and X² are OH, and X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X¹ and X⁵ are OH, and X², X³, X⁴, X⁶, X⁷, and X⁸ are H;

X¹ and X⁷ are OH, and X², X³, X⁴, X⁵, X⁶, and X⁸ are H;

X¹ and X⁸ are OH, and X², X³, X⁴, X⁵, X⁶, and X⁷ are H;

X² and X³ are OH, and X¹, X⁴, X⁵, X⁶, X⁷, and X⁸ are H;

X² and X⁴ are OH, and X¹, X³, X⁵, X⁶, X⁷, and X⁸ are H;

X² and X⁵ are OH, and X¹, X³, X⁴, X⁶, X⁷, and X⁸ are H;

X² and X⁶ are OH, and X¹, X³, X⁴, X⁵, X⁷, and X⁸ are H (urolithin A);

X² and X⁷ are OH, and X¹, X³, X⁴, X⁵, X⁶, and X⁸ are H;

X³ and X⁴ are OH, and X¹, X², X⁵, X⁶, X⁷, and X⁸ are H;

X³ and X⁵ are OH, and X¹, X², X⁴, X⁶, X⁷, and X⁸ are H;

X³ and X⁶ are OH, and X¹, X², X⁴, X⁵, X⁷, and X⁸ are H;

X⁵ and X⁶ are OH, and X¹, X², X³, X⁴, X⁷, and X⁸ are H;

X⁵ and X⁸ are OH, and X¹, X², X³, X⁴, X⁶, and X⁷ are H;

X⁶ and X⁷ are OH, and X¹, X², X³, X⁴, X⁵, and X⁸ are H;

X¹, X², and X⁵ are OH, and X³, X⁴, X⁶, X⁷, and X⁸ are H;

X¹, X², and X⁶ are OH, and X³, X⁴, X⁵, X⁷, and X⁸ are H;

X¹, X⁵, and X⁸ are OH, and X², X³, X⁴, X⁶, and X⁷ are H;

X², X⁴, and X⁶ are OH, and X¹, X³, X⁵, X⁷, and X⁸ are H;

X², X⁴, and X⁷ are OH, and X¹, X³, X⁵, X⁶, and X⁸ are H;

X², X⁶, and X⁷ are OH, and X¹, X³, X⁴, X⁵, and X⁸ are H (urolithin C);

X², X⁶, and X⁸ are OH, and X¹, X³, X⁴, X⁵, and X⁷ are H;

X², X⁷, and X⁸ are OH, and X¹, X³, X⁴, X⁵, and X⁶ are H;

X¹, X², X⁵, and X⁶ are OH, and X³, X⁴, X⁷, and X⁸ are H;

X¹, X², X⁵, and X⁷ are OH, and X³, X⁴, X⁶, and X⁸ are H;

X¹, X², X⁶, and X⁷ are OH, and X³, X⁴, X⁵, and X⁸ are H (urolithin D);

X¹, X⁶, X⁷, and X⁸ are OH, and X², X³, X⁴, and X⁵ are H;

X², X³, X⁶, and X⁷ are OH, and X¹, X⁴, X⁵, and X⁸ are H;

X², X⁴, X⁵, and X⁸ are OH, and X¹, X³, X⁶, and X⁷ are H;

X², X⁴, X⁶, and X⁷ are OH, and X¹, X³, X⁵, and X⁸ are H;

X¹, X², X⁴, X⁵, and X⁷ are OH, and X³, X⁶, and X⁸ are H;

X¹, X², X⁶, X⁷, and X⁸ are OH, and X³, X⁴, and X⁵ are H; or

X¹, X², X³, X⁶, X⁷, and X⁸ are OH, and X⁴ and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least two of X¹, X², X³, X⁴, X⁵,X⁶, X⁷, and X⁸ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least three of X¹, X², X³, X⁴, X⁵,X⁶, X⁷, and X⁸ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least four of X¹, X², X³, X⁴, X⁵,X⁶, X⁷, and X⁸ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least five of X¹, X², X³, X⁴, X⁵,X⁶, X⁷, and X⁸ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least six of X¹, X², X³, X⁴, X⁵,X⁶, X⁷, and X⁸ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least seven of X¹, X², X³, X⁴, X⁵,X⁶, X⁷, and X⁸ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ areOH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹ and X³ are OH; and X², X⁴, X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹ and X⁴ are OH; and X², X³, X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹ and X⁶ are OH; and X², X³, X⁴, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X² and X⁸ are OH; and X¹, X³, X⁴, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³ and X⁷ are OH; and X¹, X², X⁴, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³ and X⁸ are OH; and X¹, X², X⁴, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴ and X⁵ are OH; and X¹, X², X³, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴ and X⁶ are OH; and X¹, X², X³, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴ and X⁷ are OH; and X¹, X², X³, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴ and X⁸ are OH; and X¹, X², X³, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁵ and X⁷ are OH; and X¹, X², X³, X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁶ and X⁸ are OH; and X¹, X², X³, X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁷ and X⁸ are OH; and X¹, X², X³, X⁴,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², and X³ are OH; and X⁴, X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², and X⁴ are OH; and X³, X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², and X⁷ are OH; and X³, X⁴, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², and X⁸ are OH; and X³, X⁴, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, and X⁴ are OH; and X², X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, and X⁵ are OH; and X², X⁴, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, and X⁶ are OH; and X², X⁴, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, and X⁷ are OH; and X², X⁴, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, and X⁸ are OH; and X², X⁴, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, and X⁵ are OH; and X², X³, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, and X⁶ are OH; and X², X³, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, and X⁷ are OH; and X², X³, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, and X⁸ are OH; and X², X³, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁵, and X⁶ are OH; and X², X³, X⁴,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁵, and X⁷ are OH; and X², X³, X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁶, and X⁷ are OH; and X², X³, X⁴,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁶, and X⁸ are OH; and X², X³, X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁷, and X⁸ are OH; and X², X³, X⁴,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, and X⁴ are OH; and X¹, X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, and X⁵ are OH; and X¹, X⁴, X⁶,X⁷ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, and X⁶ are OH; and X¹, X⁴, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, and X⁷ are OH; and X¹, X⁴, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, and X⁸ are OH; and X¹, X⁴, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, and X⁵ are OH; and X¹, X³, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, and X⁸ are OH; and X¹, X³, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, and X⁶ are OH; and X¹, X³, X⁴,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, and X⁷ are OH; and X¹, X³, X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, and X⁸ are OH; and X¹, X³, X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, and X⁵ are OH; and X¹, X², X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, and X⁶ are OH; and X¹, X², X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, and X⁷ are OH; and X¹, X², X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, and X⁸ are OH; and X¹, X², X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, and X⁶ are OH; and X¹, X², X⁴,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, and X⁷ are OH; and X¹, X², X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, and X⁸ are OH; and X¹, X², X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁶, and X⁷ are OH; and X¹, X², X⁴,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁶, and X⁸ are OH; and X¹, X², X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁷, and X⁸ are OH; and X¹, X², X⁴,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, and X⁶ are OH; and X¹, X², X³,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, and X⁷ are OH; and X¹, X², X³,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, and X⁸ are OH; and X¹, X², X³,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁶, and X⁷ are OH; and X¹, X², X³,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁶, and X⁸ are OH; and X¹, X², X³,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁷, and X⁸ are OH; and X¹, X², X³,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁵, X⁶, and X⁷ are OH; and X¹, X², X³,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁵, X⁶, and X⁸ are OH; and X¹, X², X³,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁵, X⁷, and X⁸ are OH; and X¹, X², X³,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁶, X⁷, and X⁸ are OH; and X¹, X², X³,X⁴, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, and X⁴ are OH; and X⁵, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, and X⁵ are OH; and X⁴, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, and X⁶ are OH; and X⁴, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, and X⁷ are OH; and X⁴, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, and X⁸ are OH; and X⁴, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, and X⁵ are OH; and X³, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, and X⁶ are OH; and X³, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, and X⁷ are OH; and X³, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, and X⁸ are OH; and X³, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁵, and X⁸ are OH; and X³, X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁶, and X⁸ are OH; and X³, X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁷, and X⁸ are OH; and X³, X⁴,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, and X⁵ are OH; and X², X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, and X⁶ are OH; and X², X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, and X⁷ are OH; and X², X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, and X⁸ are OH; and X², X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, and X⁶ are OH; and X², X⁴,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, and X⁷ are OH; and X², X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, and X⁸ are OH; and X², X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁶, and X⁷ are OH; and X², X⁴,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁶, and X⁸ are OH; and X², X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁷, and X⁸ are OH; and X², X⁴,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, and X⁶ are OH; and X², X³,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, and X⁷ are OH; and X², X³,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, and X⁸ are OH; and X², X³,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁶, and X⁷ are OH; and X², X³,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁶, and X⁸ are OH; and X², X³,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁷, and X⁸ are OH; and X², X³,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁵, X⁶, and X⁷ are OH; and X², X³,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁵, X⁶, and X⁸ are OH; and X², X³,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁵, X⁷, and X⁸ are OH; and X², X³,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, and X⁵ are OH; and X¹, X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, and X⁶ are OH; and X¹, X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, and X⁷ are OH; and X¹, X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, and X⁸ are OH; and X¹, X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, and X⁶ are OH; and X¹, X⁴,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, and X⁷ are OH; and X¹, X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, and X⁸ are OH; and X¹, X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁶, and X⁸ are OH; and X¹, X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁷, and X⁸ are OH; and X¹, X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁵, and X⁶ are OH; and X¹, X³,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁵, and X⁷ are OH; and X¹, X³,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁶, and X⁸ are OH; and X¹, X³,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁷, and X⁸ are OH; and X¹, X³,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, X⁶, and X⁷ are OH; and X¹, X³,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, X⁶, and X⁸ are OH; and X¹, X³,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, X⁷, and X⁸ are OH; and X¹, X³,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁶, X⁷, and X⁸ are OH; and X¹, X³,X⁴, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, and X⁶ are OH; and X¹, X²,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, and X⁷ are OH; and X¹, X²,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, and X⁸ are OH; and X¹, X²,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁶, and X⁷ are OH; and X¹, X²,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁶, and X⁸ are OH; and X¹, X²,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁷, and X⁸ are OH; and X¹, X²,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, X⁶, and X⁷ are OH; and X¹, X²,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, X⁶, and X⁸ are OH; and X¹, X²,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, X⁷, and X⁸ are OH; and X¹, X²,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁶, X⁷, and X⁸ are OH; and X¹, X²,X⁴, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, X⁶, and X⁷ are OH; and X¹, X²,X³, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, X⁶, and X⁸ are OH; and X¹, X²,X³, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, X⁷, and X⁸ are OH; and X¹, X²,X³, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁶, X⁷, and X⁸ are OH; and X¹, X²,X³, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁵, X⁶, X⁷, and X⁸ are OH; and X¹, X²,X³, and X⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, and X⁵ are OH; and X⁶,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, and X⁶ are OH; and X⁵,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, and X⁷ are OH; and X⁵,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, and X⁸ are OH; and X⁵,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, and X⁶ are OH; and X⁴,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, and X⁷ are OH; and X⁴,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, and X⁸ are OH; and X⁴,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁶, and X⁷ are OH; and X⁴,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁶, and X⁸ are OH; and X⁴,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁷, and X⁸ are OH; and X⁴,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁵, and X⁶ are OH; and X³,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁵, and X⁸ are OH; and X³,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁶, and X⁷ are OH; and X³,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁶, and X⁸ are OH; and X³,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁷, and X⁸ are OH; and X³,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁵, X⁶, and X⁷ are OH; and X³,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁵, X⁶, and X⁸ are OH; and X³,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁵, X⁷, and X⁸ are OH; and X³,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, and X⁶ are OH; and X²,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, and X⁷ are OH; and X²,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, and X⁸ are OH; and X²,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁶, and X⁷ are OH; and X²,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁶, and X⁸ are OH; and X²,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁷, and X⁸ are OH; and X²,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, X⁶, and X⁷ are OH; and X²,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, X⁶, and X⁸ are OH; and X²,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, X⁷, and X⁸ are OH; and X²,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁶, X⁷, and X⁸ are OH; and X²,X⁴, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, X⁶, and X⁷ are OH; and X²,X³, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, X⁶, and X⁸ are OH; and X²,X³, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, X⁷, and X⁸ are OH; and X²,X³, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁶, X⁷, and X⁸ are OH; and X²,X³, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁵, X⁶, X⁷, and X⁸ are OH; and X²,X³, and X⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, and X⁶ are OH; and X¹,X⁷, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, and X⁷ are OH; and X¹,X⁶, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, and X⁸ are OH; and X¹,X⁶, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁶, and X⁷ are OH; and X¹,X⁵, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁶, and X⁸ are OH; and X¹,X⁵, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁷, and X⁸ are OH; and X¹,X⁵, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, X⁶, and X⁷ are OH; and X¹,X⁴, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, X⁶, and X⁸ are OH; and X¹,X⁴, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, X⁷, and X⁸ are OH; and X¹,X⁴, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁶, X⁷, and X⁸ are OH; and X¹,X⁴, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁵, X⁶, and X⁷ are OH; and X¹,X³, and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁵, X⁶, and X⁸ are OH; and X¹,X³, and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁵, X⁷, and X⁸ are OH; and X¹,X³, and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁶, X⁷, and X⁸ are OH; and X¹,X³, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁵, X⁶, X⁷, and X⁸ are OH; and X¹,X³, and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, X⁶, and X⁷ are OH; and X¹,X², and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, X⁶, and X⁸ are OH; and X¹,X², and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, X⁷, and X⁸ are OH; and X¹,X², and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁶, X⁷, and X⁸ are OH; and X¹,X², and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁵, X⁶, X⁷, and X⁸ are OH; and X¹,X², and X⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁴, X⁵, X⁶, X⁷, and X⁸ are OH; and X¹,X², and X³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, and X⁶ are OH; andX⁷ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, and X⁷ are OH; andX⁶ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, and X⁸ are OH; andX⁶ and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁶, and X⁷ are OH; andX⁵ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁶, and X⁸ are OH; andX⁵ and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁷, and X⁸ are OH; andX⁵ and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, X⁶, and X⁷ are OH; andX⁴ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, X⁶, and X⁸ are OH; andX⁴ and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, X⁷, and X⁸ are OH; andX⁴ and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁵, X⁶, and X⁷ are OH; andX³ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁵, X⁶, and X⁸ are OH; andX³ and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁵, X⁷, and X⁸ are OH; andX³ and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁶, X⁷, and X⁸ are OH; andX³ and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁵, X⁶, X⁷, and X⁸ are OH; andX³ and X⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, X⁶, and X⁷ are OH; andX² and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, X⁶, and X⁸ are OH; andX² and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, X⁷, and X⁸ are OH; andX² and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁶, X⁷, and X⁸ are OH; andX² and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁵, X⁶, X⁷, and X⁸ are OH; andX² and X⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X⁴, X⁵, X⁶, X⁷, and X⁸ are OH; andX² and X³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, X⁶, and X⁷ are OH; andX¹ and X⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, X⁶, and X⁸ are OH; andX¹ and X⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, X⁷, and X⁸ are OH; andX¹ and X⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁶, X⁷, and X⁸ are OH; andX¹ and X⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁵, X⁶, X⁷, and X⁸ are OH; andX¹ and X⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X⁴, X⁵, X⁶, X⁷, and X⁸ are OH; andX¹ and X³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are OH; andX¹ and X² are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, X⁶, and X⁷ are OH;and X⁸ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, X⁶, and X⁸ are OH;and X⁷ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁵, X⁷, and X⁸ are OH;and X⁶ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁴, X⁶, X⁷, and X⁸ are OH;and X⁵ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X³, X⁵, X⁶, X⁷, and X⁸ are OH;and X⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X², X⁴, X⁵, X⁶, X⁷, and X⁸ are OH;and X³ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹, X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are OH;and X² is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X², X³, X⁴, X⁵, X⁶, X⁷, and X⁸ are OH;and X¹ is H.

In certain embodiments, the invention relates to a compound of FormulaIII

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR;

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide; and

with the proviso that the compound is not a compound of Formula IIIwherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R¹ is OR, and R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R² is OR, and R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R³ is OR, and R¹, R², R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R⁴ is OR, and R¹, R², R³, R⁵, R⁶, R⁷, and R⁸ are H;

R⁵ is OR, and R¹, R², R³, R⁴, R⁶, R⁷, and R⁸ are H;

R⁶ is OR, and R¹, R², R³, R⁴, R⁵, R⁷, and R⁸ are H;

R⁷ is OR, and R¹, R², R³, R⁴, R⁵, R⁶, and R⁸ are H;

R⁸ is OR, and R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are H;

R¹ and R² are OR, and R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R¹ and R⁵ are OR, and R², R³, R⁴, R⁶, R⁷, and R⁸ are H;

R¹ and R⁷ are OR, and R², R³, R⁴, R⁵, R⁶, and R⁸ are H;

R¹ and R⁸ are OR, and R², R³, R⁴, R⁵, R⁶, and R⁷ are H;

R² and R³ are OR, and R¹, R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R² and R⁴ are OR, and R¹, R³, R⁵, R⁶, R⁷, and R⁸ are H;

R² and R⁵ are OR, and R¹, R³, R⁴, R⁶, R⁷, and R⁸ are H;

R² and R⁶ are OR, and R¹, R³, R⁴, R⁵, R⁷, and R⁸ are H;

R² and R⁷ are OR, and R¹, R³, R⁴, R⁵, R⁶, and R⁸ are H;

R² and R⁸ are OR, and R¹, R³, R⁴, R⁵, R⁶, and R⁷ are H;

R³ and R⁴ are OR, and R¹, R², R⁵, R⁶, R⁷, and R⁸ are H;

R³ and R⁵ are OR, and R¹, R², R⁴, R⁶, R⁷, and R⁸ are H;

R³ and R⁶ are OR, and R¹, R², R⁴, R⁵, R⁷, and R⁸ are H;

R³ and R⁷ are OR, and R¹, R², R⁴, R⁵, R⁶, and R⁸ are H;

R³ and R⁸ are OR, and R¹, R², R⁴, R⁵, R⁶, and R⁷ are H;

R⁴ and R⁸ are OR, and R¹, R², R³, R⁵, R⁶, and R⁷ are H;

R⁵ and R⁶ are OR, and R¹, R², R³, R⁴, R⁷, and R⁸ are H;

R⁵ and R⁷ are OR, and R¹, R², R³, R⁴, R⁶, and R⁸ are H;

R⁵ and R⁸ are OR, and R¹, R², R³, R⁴, R⁶, and R⁷ are H;

R⁶ and R⁷ are OR, and R¹, R², R³, R⁴, R⁵, and R⁸ are H;

R⁶ and R⁸ are OR, and R¹, R², R³, R⁴, R⁵, and R⁷ are H;

R¹, R², and R³ are OR, and R⁴, R⁵, R⁶, R⁷, and R⁸ are H;

R¹, R², and R⁵ are OR, and R³, R⁴, R⁶, R⁷, and R⁸ are H;

R¹, R², and R⁶ are OR, and R³, R⁴, R⁵, R⁷, and R⁸ are H;

R¹, R², and R⁸ are OR, and R³, R⁴, R⁵, R⁶, and R⁷ are H;

R¹, R⁵, and R⁸ are OR, and R², R³, R⁴, R⁶, and R⁷ are H;

R¹, R⁷, and R⁸ are OR, and R², R³, R⁴, R⁵, and R⁶ are H;

R², R³, and R⁴ are OR, and R¹, R⁵, R⁶, R⁷, and R⁸ are H;

R², R⁴, and R⁶ are OR, and R¹, R³, R⁵, R⁷, and R⁸ are H;

R², R⁴, and R⁷ are OR, and R¹, R³, R⁵, R⁶, and R⁸ are H;

R², R⁵, and R⁸ are OR, and R¹, R³, R⁴, R⁶, and R⁷ are H;

R², R⁶, and R⁷ are OR, and R¹, R³, R⁴, R⁵, and R⁸ are H;

R², R⁶, and R⁸ are OR, and R¹, R³, R⁴, R⁵, and R⁷ are H;

R², R⁷, and R⁸ are OR, and R¹, R³, R⁴, R⁵, and R⁶ are H;

R³, R⁵, and R⁸ are OR, and R¹, R², R⁴, R⁶, and R⁷ are H;

R³, R⁷, and R⁸ are OR, and R¹, R², R⁴, R⁵, and R⁶ are H;

R⁶, R⁷, and R⁸ are OR, and R¹, R², R³, R⁴, and R⁵ are H;

R¹, R², R⁵, and R⁶ are OR, and R³, R⁴, R⁷, and R⁸ are H;

R¹, R², R⁵, and R⁷ are OR, and R³, R⁴, R⁶, and R⁸ are H;

R¹, R², R⁶, and R⁷ are OR, and R³, R⁴, R⁵, and R⁸ are H;

R¹, R⁶, R⁷, and R⁸ are OR, and R², R³, R⁴, and R⁵ are H;

R², R³, R⁴, and R⁶ are OR, and R¹, R⁵, R⁷, and R⁸ are H;

R², R³, R⁵, and R⁷ are OR, and R¹, R⁴, R⁶, and R⁸ are H;

R², R³, R⁶, and R⁷ are OR, and R¹, R⁴, R⁵, and R⁸ are H;

R², R⁴, R⁵, and R⁸ are OR, and R¹, R³, R⁶, and R⁷ are H;

R², R⁴, R⁶, and R⁷ are OR, and R¹, R³, R⁵, and R⁸ are H;

R², R⁵, R⁶, and R⁷ are OR, and R¹, R³, R⁴, and R⁸ are H;

R², R⁶, R⁷, and R⁸ are OR, and R¹, R³, R⁴, and R⁵ are H;

R¹, R², R⁴, R⁵, and R⁷ are OR, and R³, R⁶, and R⁸ are H;

R¹, R², R⁶, R⁷, and R⁸ are OR, and R³, R⁴, and R⁵ are H;

R², R³, R⁴, R⁵, and R⁷ are OR, and R¹, R⁶, and R⁸ are H;

R², R³, R⁶, R⁷, and R⁸ are OR, and R¹, R⁴, and R⁵ are H;

R², R⁴, R⁶, R⁷, and R⁸ are OR, and R¹, R³, and R⁵ are H;

R², R⁵, R⁶, R⁷, and R⁸ are OR, and R¹, R³, and R⁴ are H;

R¹, R², R³, R⁶, R⁷, and R⁸ are OR, and R⁴ and R⁵ are H;

R², R³, R⁴, R⁶, R⁷, and R⁸ are OR, and R¹ and R⁵ are H; and

R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR, and R¹ and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least two of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least three of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least four of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least five of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least six of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least seven of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areOR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹ and R³ are OR; and R², R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹ and R⁴ are OR; and R², R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹ and R⁶ are OR; and R², R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴ and R⁵ are OR; and R¹, R², R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴ and R⁶ are OR; and R¹, R², R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴ and R⁷ are OR; and R¹, R², R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁷ and R⁸ are OR; and R¹, R², R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², and R⁴ are OR; and R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², and R⁷ are OR; and R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, and R⁴ are OR; and R², R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, and R⁵ are OR; and R², R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, and R⁶ are OR; and R², R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, and R⁷ are OR; and R², R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, and R⁸ are OR; and R², R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, and R⁵ are OR; and R², R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, and R⁶ are OR; and R², R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, and R⁷ are OR; and R², R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, and R⁸ are OR; and R², R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁵, and R⁶ are OR; and R², R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁵, and R⁷ are OR; and R², R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁶, and R⁷ are OR; and R², R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁶, and R⁸ are OR; and R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, and R⁵ are OR; and R¹, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, and R⁶ are OR; and R¹, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, and R⁷ are OR; and R¹, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, and R⁸ are OR; and R¹, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, and R⁵ are OR; and R¹, R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, and R⁸ are OR; and R¹, R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁵, and R⁶ are OR; and R¹, R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁵, and R⁷ are OR; and R¹, R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, and R⁵ are OR; and R¹, R², R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, and R⁶ are OR; and R¹, R², R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, and R⁷ are OR; and R¹, R², R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, and R⁸ are OR; and R¹, R², R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁵, and R⁶ are OR; and R¹, R², R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁵, and R⁷ are OR; and R¹, R², R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁶, and R⁷ are OR; and R¹, R², R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁶, and R⁸ are OR; and R¹, R², R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, and R⁶ are OR; and R¹, R², R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, and R⁷ are OR; and R¹, R², R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, and R⁸ are OR; and R¹, R², R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁶, and R⁷ are OR; and R¹, R², R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁶, and R⁸ are OR; and R¹, R², R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁷, and R⁸ are OR; and R¹, R², R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁵, R⁶, and R⁷ are OR; and R¹, R², R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁵, R⁶, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁵, R⁷, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, and R⁴ are OR; and R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, and R⁵ are OR; and R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, and R⁶ are OR; and R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, and R⁷ are OR; and R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, and R⁸ are OR; and R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, and R⁵ are OR; and R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, and R⁶ are OR; and R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, and R⁷ are OR; and R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, and R⁸ are OR; and R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁵, and R⁸ are OR; and R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁶, and R⁸ are OR; and R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁷, and R⁸ are OR; and R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, and R⁵ are OR; and R², R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, and R⁶ are OR; and R², R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, and R⁷ are OR; and R², R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, and R⁸ are OR; and R², R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, and R⁶ are OR; and R², R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, and R⁷ are OR; and R², R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, and R⁸ are OR; and R², R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁶, and R⁷ are OR; and R², R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁶, and R⁸ are OR; and R², R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁷, and R⁸ are OR; and R², R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, and R⁶ are OR; and R², R³, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁵, and R⁷ are OR; and R², R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, and R⁸ are OR; and R², R³, R⁶,and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁶, and R⁷ are OR; and R², R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁶, and R⁸ are OR; and R², R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁷, and R⁸ are OR; and R², R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁵, R⁶, and R⁷ are OR; and R², R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁵, R⁶, and R⁸ are OR; and R², R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁵, R⁷, and R⁸ are OR; and R², R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, and R⁵ are OR; and R¹, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, and R⁷ are OR; and R¹, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, and R⁸ are OR; and R¹, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁵, and R⁶ are OR; and R¹, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁵, and R⁸ are OR; and R¹, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁶, and R⁸ are OR; and R¹, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁷, and R⁸ are OR; and R¹, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁵, and R⁶ are OR; and R¹, R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁵, and R⁷ are OR; and R¹, R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁶, and R⁸ are OR; and R¹, R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁷, and R⁸ are OR; and R¹, R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁵, R⁶, and R⁸ are OR; and R¹, R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁵, R⁷, and R⁸ are OR; and R¹, R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, and R⁶ are OR; and R¹, R²,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, and R⁷ are OR; and R¹, R²,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, and R⁸ are OR; and R¹, R²,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁶, and R⁷ are OR; and R¹, R²,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁶, and R⁸ are OR; and R¹, R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁷, and R⁸ are OR; and R¹, R²,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁵, R⁶, and R⁷ are OR; and R¹, R²,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁵, R⁶, and R⁸ are OR; and R¹, R²,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁵, R⁷, and R⁸ are OR; and R¹, R²,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, R⁶, and R⁷ are OR; and R¹, R²,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, R⁶, and R⁸ are OR; and R¹, R²,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁵, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, and R⁵ are OR; and R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, and R⁶ are OR; and R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, and R⁷ are OR; and R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, and R⁸ are OR; and R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, and R⁶ are OR; and R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, and R⁷ are OR; and R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, and R⁸ are OR; and R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁶, and R⁷ are OR; and R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁶, and R⁸ are OR; and R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁷, and R⁸ are OR; and R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁵, and R⁶ are OR; and R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁵, and R⁸ are OR; and R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁶, and R⁷ are OR; and R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁶, and R⁸ are OR; and R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁷, and R⁸ are OR; and R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁵, R⁶, and R⁷ are OR; and R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁵, R⁶, and R⁸ are OR; and R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁵, R⁷, and R⁸ are OR; and R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, and R⁶ are OR; and R²,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, and R⁷ are OR; and R²,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, and R⁸ are OR; and R²,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁶, and R⁷ are OR; and R²,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁶, and R⁸ are OR; and R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁷, and R⁸ are OR; and R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, R⁶, and R⁷ are OR; and R²,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, R⁶, and R⁸ are OR; and R²,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, R⁷, and R⁸ are OR; and R²,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁶, R⁷, and R⁸ are OR; and R²,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁵, R⁶, and R⁷ are OR; and R²,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁵, R⁶, and R⁸ are OR; and R²,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁵, R⁷, and R⁸ are OR; and R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁶, R⁷, and R⁸ are OR; and R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁵, R⁶, R⁷, and R⁸ are OR; and R²,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁵, and R⁶ are OR; and R¹,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁵, and R⁸ are OR; and R¹,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁶, and R⁷ are OR; and R¹,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁶, and R⁸ are OR; and R¹,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁷, and R⁸ are OR; and R¹,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁵, R⁶, and R⁷ are OR; and R¹,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁵, R⁶, and R⁸ are OR; and R¹,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁵, R⁷, and R⁸ are OR; and R¹,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁵, R⁶, and R⁷ are OR; and R¹,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁵, R⁶, and R⁸ are OR; and R¹,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R⁴, R⁵, R⁷, and R⁸ are OR; and R¹,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, R⁶, and R⁷ are OR; and R¹,R², and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, R⁶, and R⁸ are OR; and R¹,R², and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, R⁷, and R⁸ are OR; and R¹,R², and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, and R⁶ are OR; andR⁷ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, and R⁷ are OR; andR⁶ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, and R⁸ are OR; andR⁶ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁶, and R⁷ are OR; andR⁵ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁶, and R⁸ are OR; andR⁵ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁷, and R⁸ are OR; andR⁵ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, R⁶, and R⁷ are OR; andR⁴ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, R⁶, and R⁸ are OR; andR⁴ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, R⁷, and R⁸ are OR; andR⁴ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁵, R⁶, and R⁷ are OR; andR³ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁵, R⁶, and R⁸ are OR; andR³ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁵, R⁷, and R⁸ are OR; andR³ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁶, R⁷, and R⁸ are OR; andR³ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁵, R⁶, R⁷, and R⁸ are OR; andR³ and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, R⁶, and R⁷ are OR; andR² and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, R⁶, and R⁸ are OR; andR² and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, R⁷, and R⁸ are OR; andR² and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁶, R⁷, and R⁸ are OR; andR² and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁵, R⁶, R⁷, and R⁸ are OR; andR² and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR² and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁵, R⁶, and R⁷ are OR; andR¹ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁵, R⁶, and R⁸ are OR; andR¹ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁵, R⁷, and R⁸ are OR; andR¹ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R² are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are OR;and R⁸ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, R⁶, and R⁸ are OR;and R⁷ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁵, R⁷, and R⁸ are OR;and R⁶ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁴, R⁶, R⁷, and R⁸ are OR;and R⁵ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R³, R⁵, R⁶, R⁷, and R⁸ are OR;and R⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R³ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R² is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R¹ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R is H.

As used herein, the term “urolithin precursor” refers to ellagic acidand any ellagitannin capable of being converted to one or moreurolithins following administration to an animal. In certainembodiments, a “urolithin precursor” is a compound of Formula IV:

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a compound of Formula V

wherein

X⁹, X¹⁰, X¹¹, X¹², X¹³, and X¹⁴ are independently selected from thegroup consisting of H and OH; and

with the proviso that the compound is not a compound of Formula Vwherein

X⁹, X¹⁰, X¹¹, X¹², X¹³, and X¹⁴ are H;

X¹⁰ is OH, and X⁹, X¹¹, X¹², X¹³, and X¹⁴ are H;

X⁹ and X¹² are OH, and X¹⁰, X¹¹, X¹³, and X¹⁴ are H;

X⁹ and X¹³ are OH, and X¹⁰, X¹¹, X¹², and X¹⁴ are H;

X⁹ and X¹⁴ are OH, and X¹⁰, X¹¹, X¹², and X¹³ are H;

X¹⁰ and X¹³ are OH, and X⁹, X¹¹, X¹², and X¹⁴ are H;

X¹⁰, X¹¹, and X¹³ are OH, and X⁹, X¹², and X¹⁴ are H;

X⁹, X¹⁰, X¹², and X¹⁴ are OH, and X¹¹ and X¹³ are H;

X⁹, X¹⁰, X¹³, and X¹⁴ are OH, and X¹¹ and X¹² are H (ellagic acid);

X⁹, X¹⁰, X¹¹, X¹³, and X¹⁴ are OH, and X¹² is H; and

X⁹, X¹⁰, X¹¹, X¹², X¹³, and X¹⁴ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least one of X⁹, X¹⁰, X¹¹, X¹²,X¹³, and X¹⁴ is OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least two of X⁹, X¹⁰, X¹¹, X¹²,X¹³, and X¹⁴ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least three of X⁹, X¹⁰, X¹¹, X¹²,X¹³, and X¹⁴ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least four of X⁹, X¹⁰, X¹¹, X¹²,X¹³, and X¹⁴ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein five of X⁹, X¹⁰, X¹¹, X¹², X¹³, andX¹⁴ are OH.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹ is OH; and X¹⁰, X¹¹, X¹², X¹³, andX¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹¹ is OH; and X⁹, X¹⁰, X¹², X¹³, andX¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹ and X¹⁰ are OH; and X¹¹, X¹², X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹ and X¹¹ are OH; and X¹⁰, X¹², X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹⁰ and X¹¹ are OH; and X⁹, X¹², X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹⁰ and X¹² are OH; and X⁹, X¹¹, X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹¹ and X¹² are OH; and X⁹, X¹⁰, X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, and X¹¹ are OH; and X¹², X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, and X¹² are OH; and X¹¹, X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, and X¹³ are OH; and X¹¹, X¹²,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, and X¹⁴ are OH; and X¹¹, X¹²,and X¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹¹, and X¹² are OH; and X¹⁰, X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹¹, and X¹³ are OH; and X¹⁰, X¹²,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹¹, and X¹⁴ are OH; and X¹⁰, X¹²,and X¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹², and X¹³ are OH; and X¹⁰, X¹¹and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹⁰, X¹¹, and X¹² are OH; and X⁹, X¹³,and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, X¹¹, and X¹² are OH; and X¹³and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, X¹¹, and X¹³ are OH; and X¹²and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, X¹¹, and X¹⁴ are OH; and X¹²and X¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, X¹², and X¹³ are OH; and X¹¹and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹¹, X¹², and X¹³ are OH; and X¹⁰and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹¹, X¹², and X¹⁴ are OH; and X¹⁰and X¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X¹⁰, X¹¹, X¹², and X¹³ are OH; and X⁹and X¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, X¹⁰, X¹¹, X¹², and X¹³ are OH; wherein X⁹, andX¹⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein X⁹, X¹⁰, X¹¹, X¹², and X¹⁴ are OH; andX¹³ is H.

In certain embodiments, the invention relates to a compound of FormulaVI

wherein

R⁹, R¹⁰, R¹¹; R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR;

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide; and

with the proviso that the compound is not a compound of Formula VIwherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are H;

R¹⁰ is OR, and R⁹, R¹¹, R¹², R¹³, and R¹⁴ are H;

R⁹ and R¹² are OR, and R¹⁰, R¹¹, R¹³, and R¹⁴ are H;

R⁹ and R¹³ are OR, and R¹⁰, R¹¹, R¹², and R¹⁴ are H;

R⁹ and R¹⁴ are OR, and R¹⁰, R¹¹, R¹², and R¹³ are H;

R¹⁰ and R¹³ are OR, and R⁹, R¹¹, R¹², and R¹⁴ are H;

R⁹, R¹⁰, and R¹³ are OR, and R¹¹, R¹², and R¹⁴ are H;

R⁹, R¹⁰, and R¹⁴ are OR, and R¹¹, R¹², and R¹³ are H;

R¹⁰, R¹¹, and R¹³ are OR, and R⁹, R¹², and R¹⁴ are H;

R⁹, R¹⁰, R¹², and R¹³ are OR, and R¹¹ and R¹⁴ are H;

R⁹, R¹⁰, R¹², and R¹⁴ are OR, and R¹¹ and R¹³ are H;

R⁹, R¹⁰, R¹³, and R¹⁴ are OR, and R¹¹ and R¹² are H;

R¹⁰, R¹¹, R¹², and R¹³ are OR, and R⁹ and R¹⁴ are H;

R⁹, R¹⁰, R¹¹, R¹², and R¹³ are OR, and R¹⁴ is H;

R⁹, R¹⁰, R₁₁, R¹³, and R¹⁴ are OR, and R¹² is H; and

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least one of R⁹, R¹⁰, R¹¹, R¹²,R¹³, and R¹⁴ is OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least two of R⁹, R¹⁰, R¹¹, R¹²,R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least three of R⁹, R¹⁰, R¹¹, R¹²,R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein at least four of R⁹, R¹⁰, R¹¹, R¹²,R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein five of R⁹, R¹⁰, R¹¹, R¹², R¹³, andR¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹ is OR; and R¹⁰, R¹¹, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹¹ is OR; and R⁹, R¹⁰, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹ and R¹⁰ are OR; and R¹¹, R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹ and R¹¹ are OR; and R¹⁰, R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹⁰ and R¹¹ are OR; and R⁹, R¹², R¹³,R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹⁰ and R¹² are OR; and R⁹, R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹¹ and R¹² are OR; and R⁹, R¹⁰, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹⁰, and R¹¹ are OR; and R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹⁰, and R¹² are OR; and R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹¹, and R¹² are OR; and R¹⁰, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹¹, and R¹³ are OR; and R¹⁰, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹¹, and R¹⁴ are OR; and R¹⁰, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹², and R¹³ are OR; and R¹⁰, R¹¹,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R¹⁰, R¹¹, and R¹² are OR; and R⁹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹⁰, R¹¹, and R¹² are OR; and R¹³and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹⁰, R¹¹ and R¹³ are OR; and R¹²and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹⁰, R¹¹ and R¹⁴ are OR; and R¹²and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹¹, R¹², and R¹³ are OR; and R¹⁰and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹¹, R¹², and R¹⁴ are OR; and R¹⁰and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R⁹, R¹⁰, R¹¹, R¹², and R₁₄ are OR; andR¹³ is H.

In certain embodiments, the invention relates to any one of theaforementioned compounds, wherein R is H.

In one embodiment, a urolithin precursor is punicalagin (PA). In oneembodiment, a urolithin precursor is punicalin (PB). See, for example,FIG. 2 . In one embodiment, a urolithin precursor is ellagic acid (EA).In one embodiment, a urolithin precursor is provided as an isolatedurolithin precursor, e.g., isolated from a natural food source orprepared by total synthesis. Isolated urolithin precursors are usuallypurified from natural sources or synthesized de novo; some urolithinprecursors, including EA, are commercially available from suppliers,such as Sigma Aldrich.

Also in accordance with the invention, precursors of urolithins alsoinclude natural foods containing ellagitannins and ellagic acid,especially natural foods that are rich in ellagitannins, ellagic acid,or both ellagitannins and ellagic acid. Such foods include withoutlimitation certain berries, grapes, pomegranates, rose hips, and nuts.In one embodiment, the natural food is pomegranate.

Additionally, precursors of urolithins include processed foods anddrinks prepared from such natural foods. The processed food can take anyform, including, for example, jams, jellies, preserves, pastes, spreads,juices, wines, extracts, concentrates, and the like. In one embodiment,the processed food is pomegranate juice.

In one embodiment, a urolithin precursor is provided as an extract,e.g., a fruit extract.

In one embodiment, a urolithin precursor is provided as a concentrate,e.g., a fruit concentrate or fruit juice concentrate.

In one embodiment, the urolithin precursor is an isolated urolithinprecursor.

In one embodiment, the urolithin precursor is a purified urolithinprecursor.

In one embodiment, the urolithin precursor is selected from the groupconsisting of ellagic acid, an ellagitannin, and any combinationthereof.

In one embodiment, the urolithin precursor is ellagic acid.

In one embodiment, the urolithin precursor is an ellagitannin.

In one embodiment, the ellagitannin is selected from the groupconsisting of castalagin, castalin, casuarictin, chebulagic acid,chebulinic acid, gemin D, grandinin, pedunculagin, punicalagin,punicalin, roburin A, strictinin, tellimagrandin I, tellimagrandin II,terflavin A, terflavin B, tergallagin, Lambertianin C, Sanguiin H-6,Sanguiin H-10, and vescalagin.

Ellagitannins

Among the more than 500 hydrolysable tannins hitherto characterized,ellagitannins, which produce ellagic acid upon hydrolysis, constitutethe largest group; the remaining group is gallotannins(galloylglucoses). The ellagitannins include: (1) monomericellagitannins, (2)C-glycosidic ellagitannins with an open-chain glucosecore, (3) condensates of C-glycosidic tannins with flavan-3-ol (complextannin), (4) oligomers which are produced through intermolecular C—O orC—C bonds between monomers, and (5) other ellagitannins. Unlike thecondensed tannins that are widespread throughout the plant kingdom,ellagitannins have been found only in dicotyledoneous angiosperms. Amongthe plant families rich in ellagitannins are the Myrtaceae, Lythraceae,Onagraceae, Melastomataceae, and Combretaceae. These families belong tothe order Myrtales according to the plant classification systems of NewEngler, Cronquist, and APGII (angiosperm phylogeny group).

Ellagitannins are characterized by the presence of one or morehexahydroxydiphenoyl (HHDP) unit(s) on a glucopyranose core. The HHDPgroup is biosynthetically formed through intramolecular, oxidative C—Cbond formation between neighboring galloyl groups in galloylglucoses.They are easily hydrolyzed, either enzymatically or with acid, toliberate a stable ellagic acid as the dilactone form ofhexahydroxydiphenoic acid. In addition to the HHDP group, otherconstituent acyl units in ellagitannins include a galloyl group and HHDPmetabolites such as valoneoyl, dehydrohexahydroxydiphenoyl (DHHDP), andchebuloyl groups. Referring to Table 2, variations in the number andposition of these acyl units on the glucose core provide a variety ofanalogs such as tellimagrandin I (1), and II (2), pedunculagin (6),casuarictin (7), chebulagic acid (14), and chebulinic acid (15).

TABLE 2 Representative Monomeric Ellagitannins No. Name Key Structure 1Tellimagrandin I R¹ = OH; R² = R³ = G

2 Tellimagrandin II R¹ = (β)-OG; R²= R³= G 3 Strictinin R¹ = (β)-OG; R²= R³ = H 4 Gemin D R¹ = OH; R² = H; R³ = G 5 4,6-(S)-HHDP R¹ = OH; R² =R³ = H glucose 6 Pedunculagin R¹ = OH; R² = R³ = (S)-HHDP

7 Casuarictin R¹ = (β)-OG; R² = R³ = (S)-HHDP 8 2,3-(S)-HHDP R¹ = OH; R²= R³ = H glucose 9 Punicalagin R¹ = R² = (S)-HHDP

10 Punicalin R¹ = R² = H 11 Tergallagin R¹ = R² = T 12 Terflavin A R¹ =R² = (S)-HHDP

13 Terflavin B R¹ = R³ = H 14 Chebulagic acid R¹ = R² = (R)-HHDP

15 Chebulinic acid R¹ = R² = G

Table 3 presents a representative list of natural sources of monomericellagitannins 1-15.

TABLE 3 Representative Natural Sources of Monomeric Ellagitannins 1-15Ellagitannin Monomers Plant Source Found in Myrtales Trapaceae Trapajaponica 1, 4, 6, 7 Melastomataceae 6, 7 Bredia tuberculata 3, 7Heterocentron roseum 3, 7 Melastoma malabathricum 3, 6, 7 M. normale 3,6, 7 Tibouchina semidecandra 6, 7, 8 Myrtaceae Callistemon lanceolatus4, 6, 8 Eucalyptus alba 1, 4, 6, 8 E. consideniana 1, 3, 4, 6 E.globulus 1 E. rostrata 1 E. viminalis 1, 2, 4, 6 Myrtus communis 1, 2Pimenta dioica 2, 3, 5, 6 Syzygium aqueum 2, 6, 7 S. aromaticum 1, 2, 3,4, 7 Onagraceae Epilobium angustifolium 1, 3, 4, 6 Oenotheraerythrosepala 1, 4 O. laciniata 1 O. tetraptera 1, 2, 4 CombretaceaeCombretum glutinosum 8, 9, 10 C. molle 9, 10 Quisqualis indica 1, 2, 6,8, 9, 10 Terminalia arborea 8, 9, 10, 14, 15 T. arjuna 8, 9, 10 T.brachystemma 9 T. calamansanai 1, 2, 8, 9, 10 T. catappa 1, 8, 9, 10,11, 12, 13, 14, 15 T. chebula 9, 10, 12, 13, 14, 15 T. citrina 9, 14 T.macroptera 8, 9, 12, 13 T. myriocarpa 8, 9 T. triflora 10 PunicaceaePunica granatum 1, 3, 6, 8, 9, 10

C-Glycosidic ellagitannins have been found in many plant families,including Lythraceae, Myrtaceae, Combretaceae, Melastomataceae, andPunicaceae, as well as Fagaceae, Betulaceae, Casuarinaceae, Rosaceae,Theaceae, and Elaeagnaceae. They are categorized into two types:castalagin-type, which contain a flavogalloyl unit participating in theC-glucosidic linkage, such as castalagin (16) and its C-1 epimer,vescalagin (18), and casuarinin-type, which contain an HHDP unit, suchas casuarinin (20) and stachyurin (21).

TABLE 4 Representative C-Glycosidic Ellagitannins^(a) No. Name KeyStructure 16 Castalagin R¹ = H; R² = OH; R³ = R⁴ = (S)-HHDP

17 Castalin R¹ = R³ = R⁴ = H; R² = OH 18 Vescalagin R¹ = OH; R² =H; R³ =R⁴ = (S)-HHDP 19 Grandinin R¹ = L; R² = H; R³ = R⁴ = (S)-HHDP 20Casuarinin R¹ = H; R² = OH; R³ = G

21 Stachyurin R¹ = OH; R² = H; R³ = G 22 Casuariin R¹ = R³ = H; R² = OH23 5- R¹ = OH; R² = R³ = H desgalloystachyurin 29 Lagerstroemin R¹ = H;R² = OH; R³ = Val 24 Punicacortein A R¹ = R⁴ = H; R² = OH; R³ = G

25 Epi-Punicacortein R¹ = OH; R¹ = R⁴ = H; R³ = A G 26 Punicacortein BR¹ = R³ = H; R² = OH; R⁴ = G 27 Punicacortein C R¹ = H; R² = OH

28 Punicacortein D R¹ = OH; R² = H ^(a)G, (S)-HHDP, and Val have thesame meanings as in Table 2.

Table 5 presents a representative list of natural sources ofC-glycosidic ellagitannins 16-28.

TABLE 5 Representative Natural Sources of C-Glycosidic Ellagitannins16-28 C-Glycosidic Ellagitannins Family Plant species Found in MyrtalesCombretaceae Anogeissus acuminata 16, 17, 18, 19 Anogeissus leiocarpus16 Lumnitzera racemosa 16 Terminalia arjuna 16, 20, 22 Terminaliamacroptera 27 Terminalia arborea 28 Thiloa glaucocarpa 16, 18, 20, 21Lythraceae Lagerstroemia flos-reginea 16, 18, 20, 21, 22, 23, 24, 29Lagerstroemia speciosa 16, 18, 19, 29 Melastomataceae Osbeckia chinensis20, 22, 25 Tibouchina semidecandra 16, 18, 20 Myrtaceae Callistemonlanceolatus 20 Eucalyptus alba 21, 22 Eugenia grandis 16, 18 Kunzeaambigua 20 Melaleuca squarrosa 20, 21 Pimenta dioica 20, 22 16, 18,Siphoneugena densiflora 16, 20 Syzygium aqueum 16, 18, 19 Syzygiumaromaticum 20, 22 Punicaceae Punica granatum 20, 22, 25, 26, 27, 28Trapaceae Trapa japonica 20

Complex tannins (flavono-ellagitannins) are characterized by a uniqueC—C condensed structure of C-glycosidic tannins (vescalagin-type orstachyurin-type) with flavan-3-ol (catechin or epicatechin). Unlike theC-glycosidic tannins, these tannins have been found in a rather limitednumber of plant species belonging to the Combretaceae, Myrtaceae,Melastomataceae, Fagaceae, and Theaceae families. Table 6 presents arepresentative list of complex ellagitannins.

TABLE 6 Representative Complex Ellagitannins Acutissimin A Guajavin AGuajavin B Guavin A Guavin C Guavin D Malabathrin A Malabathrin EMalabathrin F Mongolicain A Mongolicain B Psidinin A Psidinin B PsidininC Stenophyllanin A

Oligomeric ellagitannins are common among many plant families, includingthe Fagaceae, Rosaceae, Coriariaceae, Onagraceae, Melastomataceae,Myrtaceae, and Lythraceae. This class of tannins is divided into threesub-groups based on structural features: (1) oligomers that contain avaloneoyl group or its equivalent, formed by intermolecular C—O bondsbetween an HHDP group and a galloyl group of a neighboring monomer; (2)macrocyclic oligomers formed by two C—O bonds; and (3)C-glycosidictannin oligomers produced by intermolecular C—C bond formation betweenC-1 of one monomer and the aromatic ring of another. Table 7 presents arepresentative list of oligomeric ellagitannins.

TABLE 7 Representative Oligomeric Ellagitannins C-glycosidicEllagitannin Dimers from Combretaceae Anogeissinin Anogeissusin AAnogeissusin B Castamollinin Other Ellagitannin Oligomers Alienanin BBenzylcation Casuarictin Casuglaunin B Cowaniin Cuphiin D1 Cuphiin D2Eugeniflorin D1 Eugeniflorin D2 Melasquanin A Melasquanin B MelasquaninC Melasquanin D Melastoflorin A Melastoflorin B Melastoflorin CMelastoflorin D Nobotanin B Nobotanin E Nobotanin F Nobotanin KOenothein A Oenothein B Oenotherin T1 Oenotherin T2 Pterocaryanin CReginin A Reginin B Reginin C Reginin D Stachyurin Woodfordin CWoodfordin D

Table 8 provides a representative list of other ellagitannins.

TABLE 8 Representative Other Ellagitannins Acalyphidin D1 AgrimoniinAscorgeraniin (Elaeocarpusin) Camelliatannin A Camelliatannin BCamelliatannin E Camelliatannin F Camelliin B Coriariin ADehydrogeraniin Eucalbanin B Eucalbanin C Euphorbin E Eurobustin CFurosinin Gemin A Geraniin Geraniinic Acid B Geraniinic Acid CHeterophylliin E Hirtellin A Hirtellin B Hirtellin C Laevigatin BLaevigatin C Laevigatin E Liquidambin Melastoflorin A Phyllanthusiin APhyllanthusiin B Phyllanthusiin C Potentillin Putranjivain ARepandusinic acid Roshenin A Roshenin B Rugosin D Rugosin E Rugosin FRugosin G Tamarixinin A Tamarixinin B Tamarixinin C Terchebin Trapanin B

Table 9 provides yet another representative list of ellagitannins,including common known natural sources for some of them.

TABLE 9 Representative Ellagitannins Name Source(s)2-O-galloyl-punicalin Casaurictin Rhu tree, Stachyrus plant Castalagin &Vecalagin Pomegranate bark Castalin Casuarictin T. japonica CasuariinBanaba tree leaves Casuarinin Banaba tree leaves Casuarinin PomegranateChebulagic acid T. chebula Chebulinic acid T. chebula CorilaginPomegranate Cornusiin A Cornusiin C Cornusiin E Epipunicacortein ABanaba tree leaves Flosin B Banaba tree leaves Gemin D T. japonicaGranatin A Pomegranate Granatin B Pomegranate Grandinin LagerstroeminBanaba tree leaves Lambertianin A Lambertianin C RaspberriesPedunculagin Pomegrante bark, and pericarp Punicacortein A PomegranatePunicacortein B Pomegranate Punicacortein C Pomegranate Punicacortein DPomegranate Punicafolin Pomegranate Punicalagin Pomegranate PunicalinPomegranate Punigluconin Pomegranate Roburin A Roburin B Roburin CRoburin D Roburin E Rubusuaviin C Tea leaves Sanguiin H-4 Muscadinegrapes Sanguiin H-5 Muscadine grapes Sanguiin H-6 Raspberries,Sanguisorba Sanguiin H-10 Sanguiin H-11 Stachyurin Banaba tree leavesStrictinin Tellimagrandin I Pomegranate Tellimigrandin II PomegranateTerchebulin Terflavin A Terflavin B Tergallagin T. catappaTerminalin/Gallagyldilacton Pomegranate

Unfortunately, for the most part ellagitannins are poorly absorbed bythe human gut. However, a number of metabolites derived fromellagitannins are absorbed by the human gut, including certainmetabolites ultimately formed in the gut by commensal microorganisms(i.e., intestinal microflora).

Ellagitannins release ellagic acid under physiological conditions invivo, and ellagic acid is then gradually metabolized by the gutmicroflora in the intestine to produce urolithin A (UA), urolithin B,urolithin C, and urolithin D. Once the metabolites are absorbed, theyare further metabolized to produce urolithin glucuronides and/orsulfates, to give a combination of metabolites secreted in the bile.

Ellagic acid is normally found in relatively low amounts in planttissues. Ellagic acid is thought to be derived from ellagitannins, whichwhen broken down form hexahydroxydiphenic acid, which spontaneouslyconverts to ellagic acid. Some additional sources of ellagic acid areshown in Table 10.

TABLE 10 Representative Sources of Ellagic Acid Fruit Quantity Acai 55.4± 1.39 mg/L fresh pulp Umbu 314 mg/100 g dry weight (commercial)Camu-camu 490 mg/100 g dry weight Cagaita 289 mg/100 g dry weight(commercial) Araçá 262 mg/100 g dry weight 218 mg/100 g dry weight(commercial) Cambuci 240 mg/100 g dry weight 512 mg/100 g dry weight(commercial) Muscadine 219 mg/100 g dry weight Grapes

Pomegranate (Punica granatum) fruits are ancient medicinal foods whichhave been used for centuries in folk medicine. They are consumed freshand as juice, which is an excellent source of ellagitannins and ellagicacid. Ellagitannins in pomegranate fruit husk and juice includepunicalin, punicalagin, corilagin, casuarinin,terminalin/gallagyldilacton, pedunculagin, tellimagrandin, granatin A,and granatin B. Other parts of the pomegranate plant contain additionalellagitannins, including punicafolin, punicacortein A, punicacortein B,punicacortein C, punicacortein D, and punigluconin. Commercial juicescontain gallagyl-type ellagitannins, including punicalagin isomers(1500-1900 mg/L), undefined hydrolyzable tannins (400-500 mg/L), andellagic acid and its glycosides (120-260 mg/L). Gil et al. (2000) J.Agric. Food Chem. 48:4581-4589. Punicalagins, ellagitannins in whichgallagic and ellagic acids are linked to a glucose molecule, areabundant in pomegranate peel. Punicalagin isomers and ellagic acidderivatives are not present in the aril juice, but during industrialjuice processing they are extracted from the husk and membranesurrounding the arils and released in large quantities into the juice.

Urolithins are metabolites of ellagic acid, punicalagin (PA), punicalin(PB), tellimagrandin (TL), and other ellagitannins. Ellagic acid (EA) isabundant in pomegranate juice. Gil et al. (2000) J. Agric. Food Chem.48:4581-4589. The ellagitannin tellimagrandin (TL) has been previouslyisolated from pomegranate and other plants. Structural formulas for UA,PA, PB, EA, and TL are presented in FIG. 2 .

As mentioned above, ellagitannins generally are not absorbed in the gut.Rather, they release EA in the gut, which is only poorly absorbed in thestomach and small intestine. EA is largely metabolized by unidentifiedbacteria in the intestinal lumen to produce urolithins. Microbialmetabolism starts in the small intestine and the first metabolitesproduced retain four phenolic hydroxyls (urolithin D, four hydroxylgroups), and these are further metabolized along the intestinal tract toremove hydroxyl units leading to urolithin C (three hydroxyls),urolithin A (two hydroxyls) and B (one hydroxyl) in the distal parts ofthe colon (FIG. 3 ). The absorbed metabolites are conjugated withglucuronic acid (one or two units), and/or methyl ethers (whenortho-dihydroxyl groupings are present). Urolithin A and B conjugatesare the main metabolites detected in plasma and urine, although sometrihydroxy derivatives (hydroxyl-UA) or EA-dimethyl ether glucuronidehave also been detected in smaller amounts. The tetrahydroxy-urolithins,trihydroxy-urolithins, and EA derivatives generally are not detected inperipheral plasma, but they are absorbed in the small intestine and theyare transported to the liver where they are further metabolized andexcreted with bile to the small intestine establishing an enterohepaticcirculation that is responsible for the relatively long life ofurolithins in plasma and urine.

In addition to natural food sources, many papers have appeared on thebiosynthesis, isolation, and biological activity of tannins, especiallyellagitannins, over the last twenty years. Access to pure ellagitanninsby isolation from natural sources may be cumbersome and yield onlyrelatively small quantities of pure natural products. See, for example,Okuda et al., (1982) Chem Pharm Bull. 30:4230-4233; Okuda et al. (1982)Chem Pharm Bull. 30:234-4236. It is therefore notable that methods fortotal synthesis of many ellagitannins are known. See, for example,Khanbabaee, K., Strategies for the synthesis of ellagitannins, In:Chemistry and Biology of Ellagitannins, Ed. S. Quideau, World ScientificPublishing, Singapore, 2009, pp. 152-202, including references citedtherein.

Methods of Increasing Autophagy, Increasing Longevity, and Treating orPreventing Diseases and Disorders Using Urolithins and Related Compounds

In certain embodiments, the invention provides methods of increasing orenhancing autophagy, both in vivo and in vitro, methods of increasinglongevity, and methods of treating or preventing various diseases andconditions using urolithins and precursors thereof. In particularembodiments, a disease or disorder treated or prevented according to thepresent invention is a disease or disorder associated with reducedautophagy, or which would benefit from increased autophagy, includingbut not limited to any of the diseases and conditions described herein.

An aspect of the invention is a method of treating or preventing adisease or condition associated with, or characterized by, reduced ordecreased autophagy, or which would benefit from increased autophagy.The method includes the step of administering to a subject in needthereof a therapeutically effective amount of a urolithin or precursorthereof. In particular embodiments, any of the urolithins or precursorsthereof described herein may be used to practice any aspect of theinvention.

As used herein, unless the context makes clear otherwise, “treat,” andsimilar words such as “treatment,” “treated,” “treating,” etc.,indicates an approach for obtaining beneficial or desired results,including clinical results. Treatment can involve optionally either thereduction or amelioration of symptoms of the disease or condition, orthe delaying of the progression of the disease or condition. In someembodiments, treatment is achieved by reducing the duration of thedisease or condition. Administration of a compound described herein may,in some embodiments, treat one or more of such symptoms.

As used herein, unless the context makes clear otherwise, “prevent,” andsimilar words such as “prevention,” “prevented,” “preventing,” etc.,indicates an approach for preventing, inhibiting, or reducing thelikelihood of the onset or recurrence of a disease or condition. It alsorefers to preventing, inhibiting, or reducing the likelihood of theoccurrence or recurrence of the symptoms of a disease or condition, oroptionally an approach for delaying the onset or recurrence of a diseaseor condition or delaying the occurrence or recurrence of the symptoms ofa disease or condition. As used herein, “prevent” and similar words alsoincludes reducing the intensity, effect, symptoms or burden of a diseaseor condition prior to onset or recurrence of the disease or condition.

An aspect of the invention is a method of increasing autophagy in acell, comprising the step of contacting a cell with an effective amountof a urolithin or a precursor thereof to increase autophagy in the cell.In particular embodiments, the cell is present within a subject, e.g., amammal. In addition, the invention includes a method of increasingautophagy in a cell, wherein the cell is present in a subject, e.g., amammal, comprising the step of providing to the subject an effectiveamount of a urolithin or a precursor thereof to increase autophagy inthe cell.

An aspect of the invention is a method of increasing autophagy in acell, comprising the step of contacting a cell with an effective amountof a urolithin, or a precursor thereof, to increase autophagy in thecell.

In particular embodiments, a urolithin is a compound having a structureset forth in Formula I, Formula II, or Formula III, including any one ofthe specific compounds of Formula II or Formula III described herein. An“effective amount” as used herein refers to an amount that is sufficientto achieve or realize a specified or desired biological effect. Forexample, an effective amount of a urolithin to increase autophagy in acell is an amount of a urolithin that is sufficient to increaseautophagy in the cell.

An increase in autophagy in a cell can be measured using any suitableassay for measuring autophagy. For example, autophagy formation can bedetermined by using the fluorescent dye monodansylcadaverine (MDC)(Sigma-Aldrich, 30432). This dye selectively labels autophagic vacuoles.Biederbick A et al. (1995) Eur. J. Cell. Biol. 66:3-14. Autophagy mayalso be determined by examining the change in the ratio of the proteinsinvolved in autophagy, such as LC3-II to LC3-I, for example, usingWestern blot analysis. With such a method, an increase in theLC3-II/LC3-I ratio in a treated cell above the baseline, untreated cellsLC3-II/LC3-I ratio, would be considered as an increase in autophagy.Examination of other protein levels such as p62 may also help in theconfirmation. For the purposes of calculating the percent (%) increasein autophagy in a cell, the ratio of LC3-II/LC3-I at baseline (B-Ratio)and the ratio of LC3-II/LC3-I during treatment (T-Ratio) may beemployed. The percent (%) increase can be determined mathematically, forexample, by the formula 100×[((T-Ratio)−(B-Ratio))/(B-Ratio)].

Autophagy is said to be increased in a cell if it is measurably greaterthan autophagy that is or would be present in an untreated or placebocontrol cell. In one embodiment autophagy is said to be increased in acell if it is greater by a statistically significant amount or degreethan autophagy that is or would be present in an untreated or placebocontrol cell. In certain embodiments, the increase in autophagy is anincrease of at least 5%, at least 10%, at least 15%, at least 20%, atleast 25%, at least 30%, at least 40%, at least 50%, at least 60%, atleast 70%, at least 80%, at least 90%, at least 100%, at least 150%, atleast 200%, at least 300%, at least 400%, at least 500%, at least 600%,at least 700%, at least 800%, at least 900%, at least 1,000% or greaterthan 1,000%, as compared to the level of autophagy present in anuntreated cell or a cell treated with a placebo.

In certain embodiments, the increase in autophagy is an increase of5-500%, 10-500%, 15-500%, 20-500%, 25-500%, 30-500%, 40-500%, 50-500%,60-500%, at least 70-500%, 80-500%, 90-500%, 100-500%, 150-500%,200-500%, 300-500%, or 400-500%, 5-1,000%, 10-500%, 15-1,000%,20-1,000%, 25-1,000%, 30-1,000%, 40-1,000%, 50-1,000%, 60-1,000%, atleast 70-1,000%, 80-1,000%, 90-1,000%, 100-1,000%, 150-1,000%,200-1,000%, 300-1,000%, 400-1,000%, 500-1,000%, 600-1,000%, 700-1,000%,800-1,000% or 900-1,000%, as compared to the level of autophagy presentin an untreated cell or a cell treated with a placebo.

In one embodiment, the autophagy is mitophagy.

In one embodiment, the urolithin is an isolated urolithin.

In one embodiment, the urolithin is a purified urolithin.

In one embodiment, the urolithin is selected from the group consistingof urolithin A, urolithin B, urolithin C, urolithin D, and anycombination thereof.

In one embodiment, the urolithin is urolithin A.

In one embodiment, the urolithin is urolithin B.

In one embodiment, the urolithin is urolithin C.

In one embodiment, the urolithin is urolithin D.

In certain embodiments, the urolithin is a compound of Formula I,Formula II, or Formula III, including any one of the specific compoundsof these formulas described herein.

In one embodiment, the urolithin precursor is an isolated urolithinprecursor.

In one embodiment, the urolithin precursor is a purified urolithinprecursor.

In certain embodiments, the urolithin precursor is a compound of FormulaIV, Formula V, or Formula VI, including any one of the specificcompounds of these formulas described herein.

In one embodiment, the urolithin precursor is selected from the groupconsisting of ellagic acid, an ellagitannin, and any combinationthereof.

In one embodiment, the urolithin precursor is ellagic acid.

In one embodiment, the urolithin precursor is an ellagitannin.

In one embodiment, the ellagitannin is selected from the groupconsisting of castalagin, castalin, casuarictin, chebulagic acid,chebulinic acid, gemin D, grandinin, pedunculagin, punicalagin,punicalin, roburin A, strictinin, tellimagrandin I, tellimagrandin II,terflavin A, terflavin B, tergallagin, Lambertianin C, Sanguiin H-6,Sanguiin H-10, and vescalagin.

In one embodiment, the cell is selected from the group consisting of:embryonic stem cells, induced pluripotent stem cells, adult stem cells,differentiated cells, blood cells, hematopoietic cells, epithelialcells, exocrine cells, endocrine cells, connective tissue cells, adiposecells, bone cells, smooth muscle cells, striated muscle cells, nervecells, sensory cells, cardiac cells, hepatic cells, gastric cells,intestinal cells, pulmonary cells, kidney cells, and germ cells. In oneembodiment, the cell is selected from the group consisting of:endothelial cells, cells of the central and peripheral nervous system(neurons (all types) and glial cells (microglia, astroglia andoligodendrocytes, Schwann cells), keratinocytes (skin cells), retinalcells, immune cells, also hair cells and follicule stem cells, andcancer stem cells. In certain embodiments, the cells are embryonic stemcells. In certain embodiments, the cells are induced pluripotent stemcells. In certain embodiments, the cells are adult stem cells. Incertain embodiments, the cells are hematopoietic stem cells. In certainembodiments the cells are cancer cells. In certain embodiments, thecells are present in an isolated organ or bloc of organs, such as anorgan or bloc of organs harvested for transplantation or maintained exvivo. In certain embodiments, the cells are present in a tissue ortissue slice. In various embodiments, the cells are contacted with theurolithin or precursor thereof in vivo, ex vivo, or in vitro.

An aspect of the invention is a method of increasing longevity in ananimal, comprising the step of administering to an animal in needthereof an effective amount of a urolithin or a precursor thereof toincrease autophagy in the animal, thereby increasing longevity of theanimal.

The term “longevity” as used herein with reference to longevity of ananimal refers to the lifespan of an individual organism. Althoughlongevity can be measured in an individual organism, it is common tomeasure and compare mean or median longevity of populations ofindividual organisms. For example, longevity may be measured andcompared as mean survival in an experimental treatment group and asuitably selected untreated or placebo control group. Longevity may alsobe considered in populations of individuals affected with a healthcondition in need of autophagy. Treatment of such individuals shallincrease their longevity as compared to their untreated counterparts. Inone embodiment, longevity is actual longevity. In one embodiment,longevity is actuarial longevity.

The term “longevity” as used herein with reference to longevity ofeukaryotic cells in vitro refers to the lifespan of an individual cell.Although longevity can be measured for an individual cell, it is commonto measure and compare mean or median longevity of populations ofindividual cells. For example, longevity may be measured and compared interms of mean survival in an experimental treatment group and a suitablyselected untreated or placebo control group. The term “longevity” alsorefers to the lifespan of a cell that is subjected to certain metabolicstresses. In one embodiment, metabolic stresses are due to nutrientdeprivation, growth factor depletion, or hypoxia. For example, treatmentof metabolically stressed cells by a urolithin or a precursor, eitherdirectly in vitro, ex vivo or in vivo by administration to a subject,would lead to an increase in the longevity of these metabolicallystressed cells and in the case of ex vivo and in vivo treatment, anincreased longevity of these metabolically stressed cells would improvethe function of tissues which they comprise.

In one embodiment, longevity is said to be increased when it is at least5 percent longer than untreated control. In various specificembodiments, longevity is said to be increased when it is at least 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, or 50 percent longer than untreated control. Incertain embodiments, longevity is increased by at least 10 percentcompared to untreated control. In certain embodiments, longevity isincreased by at least 20 percent compared to untreated control. Incertain embodiments, longevity is increased by at least 30 percentcompared to untreated control. In certain embodiments, longevity isincreased by at least 40 percent compared to untreated control. Incertain embodiments, longevity is increased by at least 50 percentcompared to untreated control.

In one embodiment, longevity is said to be increased when it is at least5 percent longer than placebo control. In various specific embodiments,longevity is said to be increased when it is at least 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28,29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46,47, 48, 49, or 50 percent longer than placebo control. In certainembodiments, longevity is increased by at least 10 percent compared toplacebo control. In certain embodiments, longevity is increased by atleast 20 percent compared to placebo control. In certain embodiments,longevity is increased by at least 30 percent compared to placebocontrol. In certain embodiments, longevity is increased by at least 40percent compared to placebo control. In certain embodiments, longevityis increased by at least 50 percent compared to placebo control.

In one embodiment, longevity is said to be increased when it is longer,by a statistically significant difference, compared to untreatedcontrol. In one embodiment, the statistical significance of thedifference is p≤0.05. In one embodiment, the statistical significance ofthe difference is p≤0.01. In one embodiment, the statisticalsignificance of the difference is p≤0.005. In one embodiment, thestatistical significance of the difference is p≤0.001.

In one embodiment, longevity is said to be increased when it is longer,by a statistically significant difference, compared to placebo control.In one embodiment, the statistical significance of the difference isp≤0.05. In one embodiment, the statistical significance of thedifference is p≤0.01. In one embodiment, the statistical significance ofthe difference is p≤0.005. In one embodiment, the statisticalsignificance of the difference is p≤0.001.

An animal is any multicellular eukaryote belonging to the kingdomAnimalia. In one embodiment an animal is an invertebrate, for example anematode (e.g., C. elegans) or a fruit fly (e.g., D. melanogaster). Inone embodiment, an animal is a vertebrate, for example a fish or amammal. In one embodiment, an animal is a mammal. In one embodiment, ananimal is a primate. In one embodiment, an animal is a human. In certainembodiments, an animal is a domestic animal, such as a dog or a cat. Incertain embodiments, an animal is a livestock, such as a horse, a cow,or a sheep.

In one embodiment, the urolithin is an isolated urolithin.

In one embodiment, the urolithin is a purified urolithin.

In one embodiment, the urolithin is selected from the group consistingof urolithin A, urolithin B, urolithin C, urolithin D, and anycombination thereof.

In one embodiment, the urolithin is urolithin A.

In one embodiment, the urolithin is urolithin B.

In one embodiment, the urolithin is urolithin C.

In one embodiment, the urolithin is urolithin D.

In certain embodiments, the urolithin is a compound of Formula I,Formula II, or Formula III, including any one of the specific compoundsof these formulas described herein.

In one embodiment, the urolithin precursor is an isolated urolithinprecursor.

In one embodiment, the urolithin precursor is a purified urolithinprecursor.

In certain embodiments, the urolithin precursor is a compound of FormulaIV, Formula V, or Formula VI, including any one of the specificcompounds of these formulas described herein.

In one embodiment, the urolithin precursor is selected from the groupconsisting of ellagic acid, an ellagitannin, and any combinationthereof.

In one embodiment, the urolithin precursor is ellagic acid.

In one embodiment, the urolithin precursor is an ellagitannin.

In one embodiment, the ellagitannin is selected from the groupconsisting of castalagin, castalin, casuarictin, chebulagic acid,chebulinic acid, gemin D, grandinin, pedunculagin, punicalagin,punicalin, roburin A, strictinin, tellimagrandin I, tellimagrandin II,terflavin A, terflavin B, tergallagin, Lambertianin C, Sanguiin H-6,Sanguiin H-10, and vescalagin.

An aspect of the invention is a method of increasing longevity ofeukaryotic cells in vitro, the method comprising the step of contactingeukaryotic cells in vitro with an effective amount of a urolithin toincrease autophagy in the cells, thereby increasing longevity of theeukaryotic cells in vitro.

In accordance with the invention, urolithins can be used as a cellculture reagent to help promote the growth and preservation of cells andtissues in culture. It is believed urolithin has the potential to keepprimary cells and tissues alive for extended periods of time, whichmeans that they could be used for a wide range of in vitro applications,including: (i) routine cell culture in research labs of cell lines,primary cells of any origin (i.e., recently isolated from humans oranimals); and (ii) tissues or organs that are maintained in culture. Forboth cells and tissues maintained in culture, this could also have invitro diagnostic applications as well as therapeutic applications, suchas: tissue expansion, protection during transport (for transplantationinto humans), for cell therapy applications using primary cells whichneed to be stored frozen, so may be part of a special solution for cellfreezing. The invention includes a method of culturing or preservingcells or tissue, comprising growing or culturing the cells or tissue ina culture medium comprising a urolithin. Cell culture media suitable forculturing and growing various cells and tissues are known in the art andcommercially available.

Urolithins and precursors thereof may also find use as positive controlswhen testing for autophagy and improved mitochondrial function in acell, tissue, or organism. For example, a urolithin can be usedseparately or included as part of a kit useful to examine mitochondrialfunction or longevity-related pathways such as the mTOR pathway.

In one embodiment, the autophagy is mitophagy.

In one embodiment, the urolithin is an isolated urolithin.

In one embodiment, the urolithin is a purified urolithin.

In one embodiment, the urolithin is selected from the group consistingof urolithin A, urolithin B, urolithin C, urolithin D, and anycombination thereof.

In one embodiment, the urolithin is urolithin A.

In one embodiment, the urolithin is urolithin B.

In one embodiment, the urolithin is urolithin C.

In one embodiment, the urolithin is urolithin D.

In certain embodiments, the urolithin is a compound of Formula I,Formula II, or Formula III, including any one of the specific compoundsof these formulas described herein.

In one embodiment, the urolithin precursor is an isolated urolithinprecursor.

In one embodiment, the urolithin precursor is a purified urolithinprecursor.

In certain embodiments, the urolithin precursor is a compound of FormulaIV, Formula V, or Formula VI, including any one of the specificcompounds of these formulas described herein.

In one embodiment, the urolithin precursor is selected from the groupconsisting of ellagic acid, an ellagitannin, and any combinationthereof.

In one embodiment, the urolithin precursor is ellagic acid.

In one embodiment, the urolithin precursor is an ellagitannin.

In one embodiment, the ellagitannin is selected from the groupconsisting of castalagin, castalin, casuarictin, chebulagic acid,chebulinic acid, gemin D, grandinin, pedunculagin, punicalagin,punicalin, roburin A, strictinin, tellimagrandin I, tellimagrandin II,terflavin A, terflavin B, tergallagin, Lambertianin C, Sanguiin H-6,Sanguiin H-10, and vescalagin.

In one embodiment, the eukaryotic cells are eukaryotic cells in primaryculture.

In one embodiment, the eukaryotic cells are part of a cell line.

In one embodiment, the eukaryotic cells are selected from the groupconsisting of: embryonic stem cells, induced pluripotent stem cells,adult stem cells, differentiated cells, blood cells, hematopoieticcells, epithelial cells, exocrine cells, endocrine cells, connectivetissue cells, adipose cells, bone cells, smooth muscle cells, striatedmuscle cells, nerve cells, sensory cells, cardiac cells, hepatic cells,gastric cells, intestinal cells, pulmonary cells, kidney cells, and germcells. In certain embodiments, the cells are embryonic stem cells. Incertain embodiments, the cells are induced pluripotent stem cells. Inone embodiment, the cell is selected from the group consisting of:endothelial cells, cells of the central and peripheral nervous system(neurons (all types) and glial cells (microglia, astroglia andoligodendrocytes, Schwann cells), keratinocytes (skin cells), retinalcells, immune cells, also hair cells and follicule stem cells, andcancer stem cells. In certain embodiments, the cells are adult stemcells. In certain embodiments, the cells are hematopoietic stem cells.In certain embodiments the cells are cancer cells.

In certain embodiments, the cell is present in an isolated tissue ororgan or portion or sample thereof. In certain embodiments, the cellsare present in an isolated organ or bloc of organs, such as an organ orbloc of organs harvested for transplantation or maintained ex vivo. Incertain embodiments, the cells are present in a tissue or tissue slice.

In particular embodiments, the cell is present in a primary culture,i.e., a “primary cell.” As used herein, “primary culture” refers tocells cultured directly from a tissue or a subject. In one embodiment, aprimary culture includes two or more types of cells. In one embodiment,a primary culture includes a single type of cell, e.g., endothelialcells. Cells in primary culture often can have only a limited number ofpassages or divisions.

In particular embodiments, the cell is present in a cell line. As usedherein, a “cell line” refers to an established, immortalized, andgenetically homogeneous population of cells derived from a eukaryoticanimal and maintained in vitro. In one embodiment, a cell line isderived from a mammal. In one embodiment, a cell line is derived from ahuman. Cell lines of many types are available from a number ofcommercial suppliers, including, for example, American Type CultureCollection (ATCC), Manassas, Va.

In certain embodiments, the invention relates to a method of increasingautophagy in a cell, comprising contacting the cell with an effectiveamount of a compound selected from the group consisting of: a compoundof Formula II, a compound of Formula III, a compound of Formula V, acompound of Formula VI, urolithin A, urolithin B, urolithin C, urolithinD, and ellagic acid, thereby increasing autophagy in the cell.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the autophagy is mitophagy.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin A.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin B.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin C.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin D.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is ellagic acid.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the cell is selected from the groupconsisting of: embryonic stem cells, induced pluripotent stem cells,adult stem cells, differentiated cells, blood cells, hematopoieticcells, epithelial cells, exocrine cells, endocrine cells, connectivetissue cells, adipose cells, bone cells, smooth muscle cells, striatedmuscle cells, nerve cells, sensory cells, cardiac cells, hepatic cells,gastric cells, intestinal cells, pulmonary cells, kidney cells, and germcells.

In certain embodiments, the invention relates to a method of increasinglongevity in an animal, comprising administering to an animal in needthereof an effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, thereby increasinglongevity of the animal.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin A.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin B.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin C.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin D.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is ellagic acid.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the animal is a mammal.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the mammal is a human.

In certain embodiments, the invention relates to a method of increasinglongevity of eukaryotic cells in vitro, comprising contacting eukaryoticcells in vitro with an effective amount of a compound selected from thegroup consisting of: a compound of Formula II, a compound of FormulaIII, a compound of Formula V, a compound of Formula VI, urolithin A,urolithin B, urolithin C, urolithin D, and ellagic acid, therebyincreasing longevity of the eukaryotic cells in vitro.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin A.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin B.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin C.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin D.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is ellagic acid.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are eukaryoticcells in primary culture.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are part of a cellline.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are cells selectedfrom the group consisting of: embryonic stem cells, induced pluripotentstem cells, adult stem cells, differentiated cells, blood cells,hematopoietic cells, epithelial cells, exocrine cells, endocrine cells,connective tissue cells, adipose cells, bone cells, smooth muscle cells,striated muscle cells, nerve cells, sensory cells, cardiac cells,hepatic cells, gastric cells, intestinal cells, pulmonary cells, kidneycells, and germ cells.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are cells selectedfrom the group consisting of: embryonic stem cells, induced pluripotentstem cells, and adult stem cells.

In certain embodiments, the invention relates to a method of increasingautophagy in a cell, comprising contacting the cell with an effectiveamount of a compound of Formula I, thereby increasing autophagy in thecell;

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the autophagy is mitophagy.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the cell is selected from the groupconsisting of: embryonic stem cells, induced pluripotent stem cells,adult stem cells, differentiated cells, blood cells, hematopoieticcells, epithelial cells, exocrine cells, endocrine cells, connectivetissue cells, adipose cells, bone cells, smooth muscle cells, striatedmuscle cells, nerve cells, sensory cells, cardiac cells, hepatic cells,gastric cells, intestinal cells, pulmonary cells, kidney cells, and germcells.

In certain embodiments, the invention relates to a method of increasinglongevity in an animal, comprising administering to an animal in needthereof an effective amount of a compound of Formula I, therebyincreasing longevity of the animal,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the animal is a mammal.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the mammal is a human.

In certain embodiments, the invention relates to a method of increasinglongevity of eukaryotic cells in vitro, comprising contacting eukaryoticcells in vitro with an effective amount of a compound of Formula I,thereby increasing longevity of the eukaryotic cells in vitro,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are eukaryoticcells in primary culture.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are part of a cellline.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are cells selectedfrom the group consisting of: embryonic stem cells, induced pluripotentstem cells, adult stem cells, differentiated cells, blood cells,hematopoietic cells, epithelial cells, exocrine cells, endocrine cells,connective tissue cells, adipose cells, bone cells, smooth muscle cells,striated muscle cells, nerve cells, sensory cells, cardiac cells,hepatic cells, gastric cells, intestinal cells, pulmonary cells, kidneycells, and germ cells.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are cells selectedfrom the group consisting of: embryonic stem cells, induced pluripotentstem cells, and adult stem cells.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areH.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least one of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ is OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least two of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least three of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least four of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least five of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least six of R¹, R², R³, R⁴, R⁵, R⁶,and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least seven of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areOR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ is OR; and R², R³, R⁴, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² is OR; and R¹, R³, R⁴, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ is OR; and R¹, R², R⁴, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ is OR; and R¹, R², R³, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ is OR; and R¹, R², R³, R⁴, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶ is OR; and R¹, R², R³, R⁴, R⁵, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁷ is OR; and R¹, R², R³, R⁴, R⁵, R⁶,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁸ is OR; and R¹, R², R³, R⁴, R⁵, R⁶,and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R² are OR; and R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R³ are OR; and R², R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁴ are OR; and R², R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁵ are OR; and R₂, R³, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁶ are OR; and R², R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁷ are OR; and R², R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁸ are OR; and R², R³, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R³ are OR; and R¹, R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁴ are OR; and R¹, R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁵ are OR; and R¹, R³, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁶ are OR; and R¹, R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁷ are OR; and R¹, R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁸ are OR; and R¹, R³, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁴ are OR; and R¹, R², R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁵ are OR; and R¹, R², R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁶ are OR; and R¹, R², R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁷ are OR; and R¹, R², R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁸ are OR; and R¹, R², R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁵ are OR; and R¹, R², R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁶ are OR; and R¹, R², R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁷ are OR; and R¹, R², R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁸ are OR; and R¹, R², R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ and R⁶ are OR; and R¹, R², R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ and R⁷ are OR; and R¹, R², R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ and R⁸ are OR; and R¹, R², R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶ and R⁷ are OR; and R¹, R², R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶ and R⁸ are OR; and R¹, R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁷ and R⁸ are OR; and R¹, R², R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R³ are OR; and R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁴ are OR; and R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁵ are OR; and R³, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁶ are OR; and R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁷ are OR; and R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁸ are OR; and R³, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁴ are OR; and R², R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁵ are OR; and R², R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁶ are OR; and R², R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁷ are OR; and R², R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁸ are OR; and R², R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁵ are OR; and R², R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁶ are OR; and R², R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁷ are OR; and R², R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁸ are OR; and R², R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, and R⁶ are OR; and R², R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, and R⁷ are OR; and R², R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, and R⁸ are OR; and R², R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁶, and R⁷ are OR; and R², R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁶, and R⁸ are OR; and R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁷, and R⁸ are OR; and R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁴ are OR; and R¹, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁵ are OR; and R¹, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁶ are OR; and R¹, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁷ are OR; and R¹, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁸ are OR; and R¹, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁵ are OR; and R¹, R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁶ are OR; and R¹, R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁷ are OR; and R¹, R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁸ are OR; and R¹, R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, and R⁶ are OR; and R¹, R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, and R⁷ are OR; and R¹, R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, and R⁸ are OR; and R¹, R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁶, and R⁷ are OR; and R¹, R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁶, and R⁸ are OR; and R¹, R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁷, and R⁸ are OR; and R¹, R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁵ are OR; and R¹, R², R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁶ are OR; and R¹, R², R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁷ are OR; and R¹, R², R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁸ are OR; and R¹, R², R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, and R⁶ are OR; and R¹, R², R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, and R⁷ are OR; and R¹, R², R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, and R⁸ are OR; and R¹, R², R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁶, and R⁷ are OR; and R¹, R², R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁶, and R⁸ are OR; and R¹, R², R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁷, and R⁸ are OR; and R¹, R², R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, and R⁶ are OR; and R¹, R², R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, and R⁷ are OR; and R¹, R², R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, and R⁸ are OR; and R¹, R², R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁶, and R⁷ are OR; and R¹, R², R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁶, and R⁸ are OR; and R¹, R², R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁷, and R⁸ are OR; and R¹, R², R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁶, and R⁷ are OR; and R¹, R², R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁶, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁷, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶, R⁷, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁴ are OR; and R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁵ are OR; and R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁶ are OR; and R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁷ are OR; and R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁸ are OR; and R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁵ are OR; and R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁶ are OR; and R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁷ are OR; and R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁸ are OR; and R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, and R⁶ are OR; and R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁷ are OR; and R³, R⁴, R⁶,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, and R⁸ are OR; and R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁶, and R⁷ are OR; and R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁶, and R⁸ are OR; and R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁷, and R⁸ are OR; and R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁵ are OR; and R², R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁶ are OR; and R², R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁷ are OR; and R², R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁸ are OR; and R², R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, and R⁶ are OR; and R², R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, and R⁷ are OR; and R², R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, and R⁸ are OR; and R², R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁶, and R⁷ are OR; and R², R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁶, and R⁸ are OR; and R², R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁷, and R⁸ are OR; and R², R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁶ are OR; and R², R³, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, and R⁷ are OR; and R², R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁸ are OR; and R², R³, R⁶,and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁶, and R⁷ are OR; and R², R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁶, and R⁸ are OR; and R², R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁷, and R⁸ are OR; and R², R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁶, and R⁷ are OR; and R², R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁶, and R⁸ are OR; and R², R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁷, and R⁸ are OR; and R², R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁶, R⁷, and R⁸ are OR; and R², R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁵ are OR; and R¹, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁶ are OR; and R¹, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁷ are OR; and R¹, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁸ are OR; and R¹, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, and R⁶ are OR; and R¹, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, and R⁷ are OR; and R¹, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, and R⁸ are OR; and R¹, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁶, and R⁷ are OR; and R¹, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁶, and R⁸ are OR; and R¹, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁷, and R⁸ are OR; and R¹, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, and R⁶ are OR; and R¹, R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, and R⁷ are OR; and R¹, R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, and R⁸ are OR; and R¹, R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁶, and R⁷ are OR; and R¹, R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁶, and R⁸ are OR; and R¹, R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁷ and R⁸ are OR; and R¹, R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁶, and R⁷ are OR; and R¹, R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁶, and R⁸ are OR; and R¹, R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁷, and R⁸ are OR; and R¹, R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁶, R⁷, and R⁸ are OR; and R¹, R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, and R⁶ are OR; and R¹, R²,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, and R⁷ are OR; and R¹, R²,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, and R⁸ are OR; and R¹, R²,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁶, and R⁷ are OR; and R¹, R²,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁶, and R⁸ are OR; and R¹, R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁷, and R⁸ are OR; and R¹, R²,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁶, and R⁷ are OR; and R¹, R²,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁶, and R⁸ are OR; and R¹, R²,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁷, and R⁸ are OR; and R¹, R²,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁶, and R⁷ are OR; and R¹, R²,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁶, and R⁸ are OR; and R¹, R²,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁵ are OR; and R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁶ are OR; and R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁷ are OR; and R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁸ are OR; and R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, and R⁶ are OR; and R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, and R⁷ are OR; and R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, and R⁸ are OR; and R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁶, and R⁷ are OR; and R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁶, and R⁸ are OR; and R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁷, and R⁸ are OR; and R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, and R⁶ are OR; and R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, and R⁷ are OR; and R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, and R⁸ are OR; and R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁶, and R⁷ are OR; and R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁶, and R⁸ are OR; and R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁷, and R⁸ are OR; and R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁶, and R⁷ are OR; and R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁶, and R⁸ are OR; and R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁷, and R⁸ are OR; and R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁶, R⁷, and R⁸ are OR; and R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, and R⁶ are OR; and R²,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, and R⁷ are OR; and R²,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, and R⁸ are OR; and R²,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁶, and R⁷ are OR; and R²,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁶, and R⁸ are OR; and R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁷, and R⁸ are OR; and R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁶, and R⁷ are OR; and R²,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁶, and R⁸ are OR; and R²,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁷, and R⁸ are OR; and R²,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁶, R⁷, and R⁸ are OR; and R²,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁶, and R⁷ are OR; and R²,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁶, and R⁸ are OR; and R²,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁷, and R⁸ are OR; and R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁶, R⁷, and R⁸ are OR; and R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁶, R⁷, and R⁸ are OR; and R²,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, and R⁶ are OR; and R¹,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, and R⁷ are OR; and R¹,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, and R⁸ are OR; and R¹,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁶, and R⁷ are OR; and R¹,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁶, and R⁸ are OR; and R¹,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁷, and R⁸ are OR; and R¹,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁶, and R⁷ are OR; and R¹,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁶, and R⁸ are OR; and R¹,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁷, and R⁸ are OR; and R¹,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁶, R⁷, and R⁸ are OR; and R¹,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁶, and R⁷ are OR; and R¹,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁶, and R⁸ are OR; and R¹,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁷, and R⁸ are OR; and R¹,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁶, R⁷, and R⁸ are OR; and R¹,R³, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁶, and R⁷ are OR; and R¹,R², and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁶, and R⁸ are OR; and R¹,R², and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁷, and R⁸ are OR; and R¹,R², and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, and R⁶ are OR; andR⁷ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, and R⁷ are OR; andR⁶ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, and R⁸ are OR; andR⁶ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁶, and R⁷ are OR; andR⁵ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁶, and R⁸ are OR; andR⁵ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁷, and R⁸ are OR; andR⁵ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁶, and R⁷ are OR; andR⁴ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁶, and R⁸ are OR; andR⁴ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁷, and R⁸ are OR; andR⁴ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁶, R⁷, and R⁸ are OR; andR⁴ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁶, and R⁷ are OR; andR³ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁶, and R⁸ are OR; andR³ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁷, and R⁸ are OR; andR³ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁶, R⁷, and R⁸ are OR; andR³ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁶, R⁷, and R⁸ are OR; andR³ and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁶, and R⁷ are OR; andR² and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁶, and R⁸ are OR; andR² and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁷, and R⁸ are OR; andR² and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁶, R⁷, and R⁸ are OR; andR² and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁶, R⁷, and R⁸ are OR; andR² and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR² and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁶, and R⁷ are OR; andR¹, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁶, and R⁸ are OR; andR¹ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁷, and R⁸ are OR; andR¹ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁶, R⁷, and R⁸ are OR; andR¹ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R² are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are OR;and R⁸ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, and R⁸ are OR;and R⁷ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁷, and R⁸ are OR;and R⁶ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁶, R⁷, and R⁸ are OR;and R⁵ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁶, R⁷, and R⁸ are OR;and R⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R³ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R² is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R¹ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R is H.

In certain embodiments, the invention relates to a method of increasingautophagy in a cell, comprising contacting the cell with an effectiveamount of a compound of Formula IV, thereby increasing autophagy in thecell;

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the autophagy is mitophagy.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the cell is selected from the groupconsisting of: embryonic stem cells, induced pluripotent stem cells,adult stem cells, differentiated cells, blood cells, hematopoieticcells, epithelial cells, exocrine cells, endocrine cells, connectivetissue cells, adipose cells, bone cells, smooth muscle cells, striatedmuscle cells, nerve cells, sensory cells, cardiac cells, hepatic cells,gastric cells, intestinal cells, pulmonary cells, kidney cells, and germcells.

In certain embodiments, the invention relates to a method of increasinglongevity in an animal, comprising administering to an animal in needthereof an effective amount of a compound of Formula IV, therebyincreasing longevity of the animal,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the animal is a mammal.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the mammal is a human.

In certain embodiments, the invention relates to a method of increasinglongevity of eukaryotic cells in vitro, comprising contacting eukaryoticcells in vitro with an effective amount of a compound of Formula IV,thereby increasing longevity of the eukaryotic cells in vitro,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are eukaryoticcells in primary culture.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are part of a cellline.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are cells selectedfrom the group consisting of: embryonic stem cells, induced pluripotentstem cells, adult stem cells, differentiated cells, blood cells,hematopoietic cells, epithelial cells, exocrine cells, endocrine cells,connective tissue cells, adipose cells, bone cells, smooth muscle cells,striated muscle cells, nerve cells, sensory cells, cardiac cells,hepatic cells, gastric cells, intestinal cells, pulmonary cells, kidneycells, and germ cells.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the eukaryotic cells are cells selectedfrom the group consisting of: embryonic stem cells, induced pluripotentstem cells, and adult stem cells.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least one of R⁹, R¹⁰, R¹¹, R¹², R¹³,and R¹⁴ is OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least two of R⁹, R¹⁰, R¹¹, R¹², R¹³,and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least three of R⁹, R¹⁰, R¹¹, R¹²,R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least four of R⁹, R¹⁰, R¹¹, R¹², R¹³,and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein five of R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ is OR; and R¹⁰, R¹¹, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ is OR; and R⁹, R¹¹, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹¹ is OR; and R⁹, R¹⁰, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹⁰ are OR; and R¹¹, R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹¹ are OR; and R¹⁰, R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹² are OR; and R¹⁰, R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹³ are OR; and R¹⁰, R¹¹, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹⁴ are OR; and R¹⁰, R¹¹, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ and R¹¹ are OR; and R⁹, R¹², R¹³,R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ and R¹² are OR; and R⁹, R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ and R¹³ are OR; and R⁹, R¹¹, R¹²,R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹¹ and R¹² are OR; and R⁹, R¹⁰, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹¹ are OR; and R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹² are OR; and R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹³ are OR; and R¹¹, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹⁴ are OR; and R¹¹, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, and R¹² are OR; and R¹⁰, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, and R¹³ are OR; and R¹⁰, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, and R¹⁴ are OR; and R¹⁰, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹², and R¹³ are OR; and R¹⁰, R¹¹,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰, R¹¹, and R¹² are OR; and R⁹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰, R¹¹, and R¹³ are OR; and R⁹, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, and R¹² are OR; and R¹³and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, and R¹³ are OR; and R¹²and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, and R¹⁴ are OR; and R¹²and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹², and R¹³ are OR; and R¹¹and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹², and R¹⁴ are OR; and R¹¹and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹³, and R¹⁴ are OR; and R¹¹and R¹² are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, R¹², and R¹³ are OR; and R¹⁰and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, R¹², and R¹⁴ are OR; and R¹⁰and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰, R¹¹, R¹², and R¹³ are OR; and R⁹and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰; R¹¹; R¹², and R¹³ are OR; andR¹⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰; R¹¹; R¹², and R¹⁴ are OR; andR¹³ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰; R¹¹; R¹³, and R¹⁴ are OR; andR¹² is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R is H.

In certain embodiments, the invention relates to a method of treating orpreventing metabolic stress, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the metabolic stress.

In certain embodiments, the invention relates to a method of treating orpreventing cardiovascular disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the cardiovascular disease.

In certain embodiments, the invention relates to a method of treating orpreventing cardiomyopathy, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the cardiomyopathy.

In certain embodiments, the invention relates to a method of improvingmuscle function, comprising administering a therapeutically effectiveamount of a compound selected from the group consisting of: a compoundof Formula II, a compound of Formula III, a compound of Formula V, acompound of Formula VI, urolithin A, urolithin B, urolithin C, urolithinD, and ellagic acid, to a subject in need thereof, thereby improvingmuscle function.

In certain embodiments, the invention relates to a method of treating orpreventing sarcopenia, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the sarcopenia.

In certain embodiments, the invention relates to a method of treating orpreventing muscle degenerative disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the muscle degenerative disease.

In certain embodiments, the invention relates to a method of treating orpreventing Duchenne muscular dystrophy, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the Duchenne muscular dystrophy.

In certain embodiments, the invention relates to a method of treating orpreventing alcoholic liver disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the alcoholic liver disease.

In certain embodiments, the invention relates to a method of treating orpreventing nonalcoholic fatty liver disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the nonalcoholic fatty liverdisease.

In certain embodiments, the invention relates to a method of treating orpreventing drug-induced liver injury, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the drug-induced liver injury.

In certain embodiments, the invention relates to a method of treating orpreventing α1-antitrypsin deficiency, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the α1-antitrypsin deficiency.

In certain embodiments, the invention relates to a method of treating orpreventing ischemia-reperfusion injury, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the ischemia/reperfusion injury.

In certain embodiments, the invention relates to a method of treating orpreventing inflammation, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the inflammation.

In certain embodiments, the invention relates to a method of treating orpreventing inflammatory bowel disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the inflammatory bowel disease.

In certain embodiments, the invention relates to a method of treating orpreventing Crohn's disease, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the Crohn's disease.

In certain embodiments, the invention relates to a method of treating orpreventing obesity, comprising administering a therapeutically effectiveamount of a compound selected from the group consisting of: a compoundof Formula II, a compound of Formula III, a compound of Formula V, acompound of Formula VI, urolithin A, urolithin B, urolithin C, urolithinD, and ellagic acid, to a subject in need thereof, thereby treating orpreventing the obesity.

In certain embodiments, the invention relates to a method of treating orpreventing metabolic syndrome, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the metabolic syndrome.

In certain embodiments, the invention relates to a method of treating orpreventing type II diabetes mellitus, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the type II diabetes mellitus.

In certain embodiments, the invention relates to a method of treating orpreventing hyperlipidemia, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the hyperlipidemia.

In certain embodiments, the invention relates to a method of treating orpreventing osteoarthritis, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the osteoarthritis.

In certain embodiments, the invention relates to a method of treating orpreventing neurodegenerative disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the neurodegenerative disease.

In certain embodiments, the invention relates to a method of treating orpreventing Alzheimer's disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the Alzheimer's disease.

In certain embodiments, the invention relates to a method of treating orpreventing Parkinson's disease, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the Parkinson's disease.

In certain embodiments, the invention relates to a method of treating orpreventing amyotrophic lateral sclerosis (ALS), comprising administeringa therapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the ALS.

In certain embodiments, the invention relates to a method of treating orpreventing cancer, comprising administering a therapeutically effectiveamount of a compound selected from the group consisting of: a compoundof Formula II, a compound of Formula III, a compound of Formula V, acompound of Formula VI, urolithin A, urolithin B, urolithin C, urolithinD, and ellagic acid, to a subject in need thereof, thereby treating orpreventing the cancer.

In certain embodiments, the invention relates to a method of treating orpreventing cognitive disorder, comprising administering atherapeutically effective amount of a compound selected from the groupconsisting of: a compound of Formula II, a compound of Formula III, acompound of Formula V, a compound of Formula VI, urolithin A, urolithinB, urolithin C, urolithin D, and ellagic acid, to a subject in needthereof, thereby treating or preventing the cognitive disorder.

In certain embodiments, the invention relates to a method of treating orpreventing mood disorder, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebytreating or preventing the mood disorder.

In certain embodiments, the invention relates to a method of treating orpreventing stress, comprising administering a therapeutically effectiveamount of a compound selected from the group consisting of: a compoundof Formula II, a compound of Formula III, a compound of Formula V, acompound of Formula VI, urolithin A, urolithin B, urolithin C, urolithinD, and ellagic acid, to a subject in need thereof, thereby treating orpreventing the stress.

In certain embodiments, the invention relates to a method of improvingactivity during aging, comprising administering a therapeuticallyeffective amount of a compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, a compound of Formula VI, urolithin A, urolithin B, urolithin C,urolithin D, and ellagic acid, to a subject in need thereof, therebyimproving activity during aging.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is a compound of Formula IIas defined herein.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is a compound of FormulaIII as defined herein.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is a compound of Formula Vas defined herein.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is a compound of Formula VIas defined herein.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin A.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin B.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin C.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is urolithin D.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein the compound is ellagic acid.

In certain embodiments, the invention relates to a method of treating orpreventing metabolic stress, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the metabolic stress,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cardiovascular disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the cardiovasculardisease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cardiomyopathy, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the cardiomyopathy,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of improvingmuscle function, comprising administering a therapeutically effectiveamount of a compound of Formula I to a subject in need thereof, therebyimproving muscle function,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing sarcopenia, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the sarcopenia,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing muscle degenerative disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the muscle degenerativedisease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Duchenne muscular dystrophy, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the Duchenne musculardystrophy,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing alcoholic liver disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the alcoholic liverdisease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing nonalcoholic fatty liver disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the nonalcoholic fattyliver disease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing drug-induced liver injury, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the drug-induced liverinjury,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing α1-antitrypsin deficiency, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the α1-antitrypsindeficiency,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing ischemia-reperfusion injury, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the ischemia/reperfusioninjury,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing inflammation, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the inflammation,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing inflammatory bowel disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the inflammatory boweldisease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Crohn's disease, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the Crohn's disease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing obesity, comprising administering a therapeutically effectiveamount of a compound of Formula I to a subject in need thereof, therebytreating or preventing the obesity,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing metabolic syndrome, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the metabolic syndrome,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing type II diabetes mellitus, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the type II diabetesmellitus,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing hyperlipidemia, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the hyperlipidemia,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing osteoarthritis, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the osteoarthritis,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing neurodegenerative disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the neurodegenerativedisease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Alzheimer's disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the Alzheimer's disease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Parkinson's disease, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the Parkinson's disease,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing amyotrophic lateral sclerosis (ALS), comprising administeringa therapeutically effective amount of a compound of Formula I to asubject in need thereof, thereby treating or preventing the ALS,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cancer, comprising administering a therapeutically effectiveamount of a compound of Formula I to a subject in need thereof, therebytreating or preventing the cancer,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cognitive disorder, comprising administering atherapeutically effective amount of a compound of Formula I to a subjectin need thereof, thereby treating or preventing the cognitive disorder,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing mood disorder, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby treating or preventing the mood disorder,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing stress, comprising administering a therapeutically effectiveamount of a compound of Formula I to a subject in need thereof, therebytreating or preventing the stress,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of improvingactivity during aging, comprising administering a therapeuticallyeffective amount of a compound of Formula I to a subject in needthereof, thereby improving activity during aging,

wherein the compound of Formula I is

wherein

R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areH.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least one of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ is OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least two of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least three of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least four of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least five of R¹, R², R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least six of R¹, R², R³, R⁴, R⁵, R⁶,and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least seven of R¹, R², R³, R⁴, R⁵,R⁶, R⁷, and R⁸ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ areOR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ is OR; and R², R³, R⁴, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² is OR; and R¹, R³, R⁴, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ is OR; and R¹, R², R⁴, R⁵, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, R³, R⁵, R⁶, R⁷, wherein R⁴ is OR; and R¹, R²,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ is OR; and R¹, R², R³, R⁴, R⁶, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶ is OR; and R¹, R², R³, R⁴, R⁵, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁷ is OR; and R¹, R², R³, R⁴, R⁵, R⁶,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁸ is OR; and R¹, R², R³, R⁴, R⁵, R⁶,and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R² are OR; and R³, R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R³ are OR; and R², R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁴ are OR; and R², R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁵ are OR; and R², R³, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁶ are OR; and R², R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁷ are OR; and R², R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹ and R⁸ are OR; and R², R³, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R³ are OR; and R¹, R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁴ are OR; and R¹, R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁵ are OR; and R¹, R³, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁶ are OR; and R¹, R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁷ are OR; and R¹, R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R² and R⁸ are OR; and R¹, R³, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁴ are OR; and R¹, R², R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁵ are OR; and R¹, R₂, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁶ are OR; and R¹, R², R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁷ are OR; and R¹, R², R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³ and R⁸ are OR; and R¹, R², R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁵ are OR; and R¹, R², R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁶ are OR; and R¹, R², R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁷ are OR; and R¹, R², R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴ and R⁸ are OR; and R¹, R², R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ and R⁶ are OR; and R¹, R², R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ and R⁷ are OR; and R¹, R², R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵ and R⁸ are OR; and R¹, R², R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶ and R⁷ are OR; and R¹, R², R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶ and R⁸ are OR; and R¹, R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁷ and R⁸ are OR; and R¹, R², R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R³ are OR; and R⁴, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁴ are OR; and R³, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁵ are OR; and R³, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁶ are OR; and R³, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁷ are OR; and R³, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁸ are OR; and R³, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁴ are OR; and R², R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁵ are OR; and R², R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁶ are OR; and R², R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁷ are OR; and R², R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, and R⁸ are OR; and R², R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁵ are OR; and R², R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁶ are OR; and R², R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁷ are OR; and R², R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁸ are OR; and R², R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, and R⁶ are OR; and R², R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, and R⁷ are OR; and R², R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, and R⁸ are OR; and R², R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁶, and R⁷ are OR; and R², R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁶, and R⁸ are OR; and R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁷, and R⁸ are OR; and R², R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁴ are OR; and R¹, R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁵ are OR; and R¹, R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁶ are OR; and R¹, R⁴, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁷ are OR; and R¹, R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, and R⁸ are OR; and R¹, R⁴, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁵ are OR; and R¹, R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁶ are OR; and R¹, R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁷ are OR; and R¹, R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, and R⁸ are OR; and R¹, R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, and R⁶ are OR; and R¹, R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, and R⁷ are OR; and R¹, R³, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, and R⁸ are OR; and R¹, R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁶, and R⁷ are OR; and R¹, R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁶, and R⁸ are OR; and R¹, R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁷, and R⁸ are OR; and R¹, R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁵ are OR; and R¹, R², R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁶ are OR; and R¹, R², R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁷ are OR; and R¹, R², R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, and R⁸ are OR; and R¹, R², R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, and R⁶ are OR; and R¹, R², R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, and R⁷ are OR; and R¹, R², R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, and R⁸ are OR; and R¹, R², R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁶, and R⁷ are OR; and R¹, R², R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁶, and R⁸ are OR; and R¹, R², R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁷, and R⁸ are OR; and R¹, R², R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, and R⁶ are OR; and R¹, R², R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, and R⁷ are OR; and R¹, R², R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, and R⁸ are OR; and R¹, R², R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁶, and R⁷ are OR; and R¹, R², R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁶, and R⁸ are OR; and R¹, R², R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁷, and R⁸ are OR; and R¹, R², R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁶, and R⁷ are OR; and R¹, R², R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁶, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁷, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁶, R⁷, and R⁸ are OR; and R¹, R², R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁴ are OR; and R⁵, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁵ are OR; and R⁴, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁶ are OR; and R⁴, R⁵, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, and R⁷ are OR; and R⁴, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁸ are OR; and R⁴, R⁵, R⁶,and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁵ are OR; and R³, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁶ are OR; and R³, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁷ are OR; and R³, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, and R⁸ are OR; and R³, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, and R⁶ are OR; and R³, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², and R⁷ are OR; and R³, R⁴, R⁶,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, and R⁸ are OR; and R³, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁶, and R⁷ are OR; and R³, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁶, and R⁸ are OR; and R³, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁷, and R⁸ are OR; and R³, R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁵ are OR; and R², R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁶ are OR; and R², R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁷ are OR; and R², R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, and R⁸ are OR; and R², R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, and R⁶ are OR; and R², R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, and R⁷ are OR; and R², R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, and R⁸ are OR; and R², R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁶, and R⁷ are OR; and R², R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁶, and R⁸ are OR; and R², R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁷, and R⁸ are OR; and R², R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁶ are OR; and R², R³, R⁷,and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, and R⁷ are OR; and R², R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, and R⁸ are OR; and R², R³, R⁶,and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁶, and R⁷ are OR; and R², R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁶, and R⁸ are OR; and R², R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, and R⁸ are OR; and R², R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁶, and R⁷ are OR; and R², R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁶, and R⁸ are OR; and R², R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁷, and R⁸ are OR; and R², R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁶, R⁷, and R⁸ are OR; and R², R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁵ are OR; and R¹, R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁶ are OR; and R¹, R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁷ are OR; and R¹, R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, and R⁸ are OR; and R¹, R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, and R⁶ are OR; and R¹, R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, and R⁷ are OR; and R¹, R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, and R⁸ are OR; and R¹, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁶, and R⁷ are OR; and R¹, R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁶, and R⁸ are OR; and R¹, R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁷, and R⁸ are OR; and R¹, R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, and R⁶ are OR; and R¹, R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, and R⁷ are OR; and R¹, R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, and R⁸ are OR; and R¹, R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁶, and R⁷ are OR; and R¹, R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁶, and R⁸ are OR; and R¹, R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁷, and R⁸ are OR; and R¹, R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁶, and R⁷ are OR; and R¹, R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁶, and R⁸ are OR; and R¹, R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁷, and R⁸ are OR; and R¹, R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁶, R⁷ and R⁸ are OR; and R¹, R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, and R⁶ are OR; and R¹, R²,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, and R⁷ are OR; and R¹, R²,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, and R⁸ are OR; and R¹, R²,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁶, and R⁷ are OR; and R¹, R²,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁶, and R⁸ are OR; and R¹, R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁷, and R⁸ are OR; and R¹, R²,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁶, and R⁷ are OR; and R¹, R²,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁶, and R⁸ are OR; and R¹, R²,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁷, and R⁸ are OR; and R¹, R²,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁶, and R⁷ are OR; and R¹, R²,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁶, and R⁸ are OR; and R¹, R²,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁵, R⁶, R⁷, and R⁸ are OR; and R¹, R²,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁵ are OR; and R⁶,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁶ are OR; and R⁵,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁷ are OR; and R⁵,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, and R⁸ are OR; and R⁵,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, and R⁶ are OR; and R⁴,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, and R⁷ are OR; and R⁴,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, and R⁸ are OR; and R⁴,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁶, and R⁷ are OR; and R⁴,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁶, and R⁸ are OR; and R⁴,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁷, and R⁸ are OR; and R⁴,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, and R⁶ are OR; and R³,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, and R⁷ are OR; and R³,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, and R⁸ are OR; and R³,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁶, and R⁷ are OR; and R³,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁶, and R⁸ are OR; and R³,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁷, and R⁸ are OR; and R³,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁶, and R⁷ are OR; and R³,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁶, and R⁸ are OR; and R³,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁷, and R⁸ are OR; and R³,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁶, R⁷, and R⁸ are OR; and R³,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, and R⁶ are OR; and R²,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, and R⁷ are OR; and R²,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, and R⁸ are OR; and R²,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁶, and R⁷ are OR; and R²,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁶, and R⁸ are OR; and R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁷, and R⁸ are OR; and R²,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁶, and R⁷ are OR; and R²,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁶, and R⁸ are OR; and R²,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁷, and R⁸ are OR; and R²,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁶, R⁷, and R⁸ are OR; and R²,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁶, and R⁷ are OR; and R²,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁶, and R⁸ are OR; and R²,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁷, and R⁸ are OR; and R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁶, R⁷, and R⁸ are OR; and R²,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁵, R⁶, R⁷, and R⁸ are OR; and R²,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, and R⁶ are OR; and R¹,R⁷, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, and R⁷ are OR; and R¹,R⁶, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, and R⁸ are OR; and R¹,R⁶, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁶, and R⁷ are OR; and R¹,R⁵, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁶, and R⁸ are OR; and R¹,R⁵, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁷, and R⁸ are OR; and R¹,R⁵, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁶, and R⁷ are OR; and R¹,R⁴, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁶, and R⁸ are OR; and R¹,R⁴, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁷, and R⁸ are OR; and R¹,R⁴, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁶, R⁷, and R⁸ are OR; and R¹,R⁴, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁶, and R⁷ are OR; and R¹,R³, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁶, and R⁸ are OR; and R¹,R³, and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁷, and R⁸ are OR; and R¹,R³, and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁶, R⁷, and R⁸ are OR; and R¹,R³, and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R³, and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁶, and R⁷ are OR; and R¹,R², and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁶, and R⁸ are OR; and R¹,R², and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁷, and R⁸ are OR; and R¹,R², and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; and R¹,R², and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, and R⁶ are OR; andR⁷ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, and R⁷ are OR; andR⁶ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, and R⁸ are OR; andR⁶ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁶, and R⁷ are OR; andR⁵ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁶, and R⁸ are OR; andR⁵ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁷, and R⁸ are OR; andR⁵ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁶, and R⁷ are OR; andR⁴ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁶, and R⁸ are OR; andR⁴ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁷, and R⁸ are OR; andR⁴ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁶, R⁷, and R⁸ are OR; andR⁴ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁶, and R⁷ are OR; andR³ and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁶, and R⁸ are OR; andR³ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁷, and R⁸ are OR; andR³ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁶, R⁷, and R⁸ are OR; andR³ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁵, R⁶, R⁷, and R⁸ are OR; andR³ and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁶, and R⁷ are OR; andR² and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁶, and R⁸ are OR; andR² and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁷, and R⁸ are OR; andR² and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁶, R⁷, and R⁸ are OR; andR² and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁵, R⁶, R⁷, and R⁸ are OR; andR² and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR² and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁶, and R⁷ are OR; andR¹, and R⁸ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁶, and R⁸ are OR; andR¹ and R⁷ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁷, and R⁸ are OR; andR¹ and R⁶ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁶, R⁷, and R⁸ are OR; andR¹ and R⁵ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR; andR¹ and R² are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, and R⁷ are OR;and R⁸ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁶, and R⁸ are OR;and R⁷ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁵, R⁷, and R⁸ are OR;and R⁶ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁴, R⁶, R⁷, and R⁸ are OR;and R⁵ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R³, R⁵, R⁶, R⁷, and R⁸ are OR;and R⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R², R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R³ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R² is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R², R³, R⁴, R⁵, R⁶, R⁷, and R⁸ are OR;and R¹ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R is H.

In certain embodiments, the invention relates to a method of treating orpreventing metabolic stress, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the metabolic stress,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cardiovascular disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the cardiovasculardisease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cardiomyopathy, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the cardiomyopathy,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of improvingmuscle function, comprising administering a therapeutically effectiveamount of a compound Formula IV to a subject in need thereof, therebyimproving muscle function,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing sarcopenia, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the sarcopenia,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing muscle degenerative disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the muscle degenerativedisease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Duchenne muscular dystrophy, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the Duchenne musculardystrophy,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing alcoholic liver disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the alcoholic liverdisease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing nonalcoholic fatty liver disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the nonalcoholic fattyliver disease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing drug-induced liver injury, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the drug-induced liverinjury,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing α1-antitrypsin deficiency, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the α1-antitrypsindeficiency,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing ischemia-reperfusion injury, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the ischemia/reperfusioninjury,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing inflammation, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the inflammation,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing inflammatory bowel disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the inflammatory boweldisease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Crohn's disease, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the Crohn's disease,

wherein the compound of Formula IV is

whereinR⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing obesity, comprising administering a therapeutically effectiveamount of a compound Formula IV to a subject in need thereof, therebytreating or preventing the obesity,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing metabolic syndrome, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the metabolic syndrome,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R₁₁, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing type II diabetes mellitus, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the type II diabetesmellitus,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing hyperlipidemia, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the hyperlipidemia,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing osteoarthritis, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the osteoarthritis,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing neurodegenerative disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the neurodegenerativedisease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Alzheimer's disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the Alzheimer's disease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing Parkinson's disease, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the Parkinson's disease,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing amyotrophic lateral sclerosis (ALS), comprising administeringa therapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the ALS,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cancer, comprising administering a therapeutically effectiveamount of a compound Formula IV to a subject in need thereof, therebytreating or preventing the cancer,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing cognitive disorder, comprising administering atherapeutically effective amount of a compound Formula IV to a subjectin need thereof, thereby treating or preventing the cognitive disorder,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing mood disorder, comprising administering a therapeuticallyeffective amount of a compound Formula IV to a subject in need thereof,thereby treating or preventing the mood disorder,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of treating orpreventing stress, comprising administering a therapeutically effectiveamount of a compound Formula IV to a subject in need thereof, therebytreating or preventing the stress,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to a method of improvingactivity during aging, comprising administering a therapeuticallyeffective amount of a compound of Formula IV to a subject in needthereof, thereby improving activity during aging,

wherein the compound of Formula IV is

wherein

R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are independently selected from thegroup consisting of H and OR; and

R is H, substituted or unsubstituted alkyl, substituted or unsubstitutedaryl, a substituted or unsubstituted monosaccharide, or a substituted orunsubstituted oligosaccharide.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least one of R⁹, R¹⁰, R¹¹, R¹², R¹³,and R¹⁴ is OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least two of R⁹, R¹⁰, R¹¹, R¹², R¹³,and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least three of R⁹, R¹⁰, R¹¹, R¹²,R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein at least four of R⁹, R¹⁰, R¹¹, R¹², R¹³,and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein five of R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹², R¹³, and R¹⁴ are OR.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ is OR; and R¹⁰, R¹¹, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ is OR; and R⁹, R¹¹, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹¹ is OR; and R⁹, R¹⁰, R¹², R¹³, andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹⁰ are OR; and R¹¹, R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹¹ are OR; and R¹⁰, R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹² are OR; and R¹⁰, R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹³ are OR; and R¹⁰, R¹¹, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹ and R¹⁴ are OR; and R¹⁰, R¹¹, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ and R¹¹ are OR; and R⁹, R¹², R¹³,R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ and R¹² are OR; and R⁹, R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰ and R¹³ are OR; and R⁹, R¹¹, R¹²,R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹¹ and R¹² are OR; and R⁹, R¹⁰, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹¹ are OR; and R¹², R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹² are OR; and R¹¹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹³ are OR; and R¹¹, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, and R¹⁴ are OR; and R¹¹, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, and R¹² are OR; and R¹⁰, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, and R¹³ are OR; and R¹⁰, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, and R¹⁴ are OR; and R¹⁰, R¹²,and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹², and R¹³ are OR; and R¹⁰, R¹¹,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰, R¹¹, and R¹² are OR; and R⁹, R¹³,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰, R¹¹, and R¹³ are OR; and R⁹, R¹²,and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, and R¹² are OR; and R¹³and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹ and R¹³ are OR; and R¹² andR¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, and R¹⁴ are OR; and R¹²and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹², and R¹³ are OR; and R¹¹and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹², and R¹⁴ are OR; and R¹¹and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹³, and R¹⁴ are OR; and R¹¹and R¹² are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, R¹², and R¹³ are OR; and R¹⁰and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹¹, R¹², and R¹⁴ are OR; and R¹⁰and R¹³ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R¹⁰, R¹¹, R¹², and R¹³ are OR; and R⁹and R¹⁴ are H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹², and R¹³ are OR; andR¹⁴ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹², and R¹⁴ are OR; andR¹³ is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R⁹, R¹⁰, R¹¹, R¹³, and R¹⁴ are OR; andR¹² is H.

In certain embodiments, the invention relates to any one of theaforementioned methods, wherein R is H.

Exemplary Compositions

In certain embodiments, the invention relates to a compositioncomprising any one of the aforementioned compounds, wherein thecomposition further comprises a pharmaceutically acceptable carrier.

In certain embodiments, the invention relates to a compositioncomprising a first compound selected from the group consisting of: acompound of Formula I, and a compound of Formula IV; and a secondcompound selected from the group consisting of rapamycin, resveratrol,metformin, and spermidine.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the first compound is urolithin A.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the first compound is urolithin B.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the first compound is urolithin C.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the first compound is urolithin D.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the first compound is ellagic acid.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the first compound is any of thespecific urolithin or urolithin precursor compounds disclosed herein.

In certain embodiments, the invention relates to any one of theaforementioned compositions, wherein the composition further comprises apharmaceutically acceptable carrier.

Compositions Comprising Urolithins and Other Compounds

An aspect of the invention is a composition, comprising a urolithin or aprecursor thereof, and one or more additional compounds that induceautophagy. In certain embodiments, the one or more additional compoundscomprises a urolithin or precursor thereof. In certain embodiments, theone or more additional compounds is selected from the group consistingof rapamycin, resveratrol, metformin, and spermidine. In a relatedembodiment, the one or more additional compounds comprise both aurolithin or precursor thereof and a compound selected from the groupconsisting of rapamycin, resveratrol, metformin, and spermidine.

In one embodiment, the urolithin is an isolated urolithin.

In one embodiment, the urolithin is a purified urolithin.

In one embodiment, the urolithin is selected from the group consistingof urolithin A, urolithin B, urolithin C, urolithin D, and anycombination thereof.

In one embodiment, the urolithin is urolithin A.

In one embodiment, the urolithin is urolithin B.

In one embodiment, the urolithin is urolithin C.

In one embodiment, the urolithin is urolithin D.

In certain embodiments, the urolithin is a compound of Formula I,Formula II, or Formula III, including any one of the specific compoundsof these formulas described herein.

In one embodiment, the urolithin precursor is an isolated urolithinprecursor.

In one embodiment, the urolithin precursor is a purified urolithinprecursor.

In certain embodiments, the urolithin precursor is a compound of FormulaIV, Formula V, or Formula VI, including any one of the specificcompounds of these formulas described herein.

In one embodiment, the urolithin precursor is selected from the groupconsisting of ellagic acid, an ellagitannin, and any combinationthereof.

In one embodiment, the urolithin precursor is ellagic acid.

In one embodiment, the urolithin precursor is an ellagitannin.

In one embodiment, the ellagitannin is selected from the groupconsisting of castalagin, castalin, casuarictin, chebulagic acid,chebulinic acid, gemin D, grandinin, pedunculagin, punicalagin,punicalin, roburin A, strictinin, tellimagrandin I, tellimagrandin II,terflavin A, terflavin B, tergallagin, Lambertianin C, Sanguiin H-6,Sanguiin H-10, and vescalagin.

In certain embodiments, the invention relates to a compositioncomprising a first compound selected from the group consisting of: acompound of Formula II, a compound of Formula III, a compound of FormulaV, and a compound of Formula VI; and a second compound selected from thegroup consisting of rapamycin, resveratrol, metformin, and spermidine.

In one embodiment, the composition further comprises a pharmaceuticallyacceptable carrier.

Autophagy is known to be strongly induced by caloric restriction (e.g.,starvation), and inhibited by the mTOR pathway. Caloric restriction,without malnutrition, has been reported to delay aging and extendlifespan in a number of species, including insects, worms, mice, rats,fish, monkeys, and even humans. See, for example, Fontana L et al.(2004) Proc Natl Acad Sci USA 101:6659-6663; Colman R J et al. (2009)Science 325:201-204. Additionally, long-term calorie restriction hasbeen reported to be highly effective in reducing the risk ofatherosclerosis in humans. Fontana L et al. (supra).

Moreover, inhibition of the mTOR pathway is known to promote autophagy,and inhibitors of the mTOR pathway are known to activate autophagy.Inhibitors of the mTOR pathway are known to include rapamycin,metformin, and resveratrol. Additional agents known to stimulateautophagy, albeit via different mechanism(s), include spermidine,clonidine, rilmenidine, tyramine, morphine, baclofen, mastoparan,propranolol, bupivacain, N-dodecyl lysinamide, tamoxifen, interferon(IFN)-gamma, trehalose and vinblastine. In one embodiment of thisaspect, the autophagy inducing compound is selected from the groupconsisting of Loperamide, Amiodarone, Niguldipine, Pimozide,Nicardipine, Penitrem A, Fluspirilene, and Trifluoerazine andpharmaceutically acceptable salts thereof.

Rapamycin((3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-9,27-dihydroxy-3-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl]-1-methylethyl]-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-23,27-epoxy-3H-pyrido[2,1-c][1,4]-oxaazacyclohentriacontine-1,5,11,28,29(4H,6H,31H)-pentone),also known as sirolimus, is a macrolide immunosuppressant agent. It wasapproved by the FDA in September 1999 and is marketed under the tradename Rapamune® by Pfizer (formerly by Wyeth). It binds to the cytosolicprotein FK-binding protein 12 (FKBP12) to form a complex that inhibitsthe mTOR pathway by directly binding the mTOR Complexi (mTORC1). In C.elegans and in yeast, rapamycin extends lifespan only under conditionsin which autophagy can be induced. Rapamycin was first shown to extendlifespan in eukaryotes in 2006. Powers et al. ((2006) Genes Dev.20:174-184) showed a dose-responsive effect of rapamycin on lifespanextension in yeast cells. Building on this and other work, in a 2009study, the lifespans of mice fed rapamycin were increased between 28 and38% from the beginning of treatment, or 9 to 14% in total increasedmaximum lifespan. Harrison D E et al. (2009) Nature 460:392-395. Ofparticular note, the treatment began in mice aged 20 months, theequivalent of 60 human years.

Metformin (N,N-Dimethylimidodicarbonimidic diamide; rosiglitazone) is athiazolidinedione compound in a class of oral antidiabetic drugs knownas the biguanide class. It is commercially available in the UnitedStates as Glucophage, Fortamet, Glumetza, and Riomet, principally foruse in the treatment of type 2 (insulin resistant) diabetes mellitus.Dowling RJO et al. (2007) Cancer Res. 67:10804-10812 reported thatmetformin inhibits the mTOR pathway in breast cancer cells.

Resveratrol (3,5,4′-trihydroxy-trans-stilbene) is a natural phenol foundin the skin of red grapes and other fruits, and it is an indirectactivator of sirtuin 1. It has also been produced by chemical synthesis.Farina A et al. (2006) Nat. Prod. Res. 20:247-252. Resveratrol isavailable as a nutritional supplement from a number of commercialsuppliers. A number of studies have reported that resveratrol extendslifespan in yeast, C. elegans, Drosophila, certain fish, and mice. Inaddition, Brito P M et al. (2009) Atherosclerosis 205:126-134 reportedthat resveratrol inhibits the mTOR pathway.

Baur J A et al. (2006) Nature 444:337-342 reported that resveratrolshifts the physiology of middle-aged mice on a high-calorie diet towardsthat of mice on a standard diet and significantly increases theirsurvival. According to Baur et al., resveratrol produces changesassociated with longer lifespan, including increased insulinsensitivity, reduced insulin-like growth factor-1 (IGF-I) levels,increased AMP-activated protein kinase (AMPK) and peroxisomeproliferator-activated receptor-gamma coactivator 1α (PGC-1α) activity,increased mitochondrial number, and improved motor function. Parametricanalysis of gene set enrichment revealed that resveratrol opposed theeffects of the high-calorie diet in 144 out of 153 significantly alteredpathways.

Pearson K J et al. (2008) Cell Metab. 8:157-168 reported thatresveratrol-fed elderly mice show a marked reduction in signs of aging,including reduced albuminuria, decreased inflammation and apoptosis inthe vascular endothelium, increased aortic elasticity, greater motorcoordination, reduced cataract formation, and preserved bone mineraldensity.

In addition to resveratrol, other small molecules have been reported toactivate sirtuins and extend lifespan in yeast, including butein(3,4,2′,4′-tetrahydroxychalcone), piceatannol(3,5,3′,4′-tetrahydroxy-trans-stilbene), isoliquiritigenin(4,2′,4′-trihydroxychalcone), fisetin (3,7,3′,4′-tetrahydroxyflavone),and quercetin (3,5,7,3′,4′-pentahydroxyflavone). Howitz K T et al.(2003) Nature 425:191-196; Wood, J G et al. (2004) Nature 430:686-689,corrigendum Nature 431:107.

The effects of various treatments on aging and longevity are commonlyassessed on invertebrates, including nematodes (C. elegans), yeast, andfruit flies (Drosophila). C. elegans, in particular, is the mostcommonly employed in vivo model for determining impact on longevity dueto its short lifespan and ease of genetic manipulation. Pharmaceuticalinterventions found to extend the lifespan of C. elegans have also beenshown to extend the lifespan of yeast and fruitflies.

Importantly, in accordance with the instant invention, urolithin A hasbeen found to be unexpectedly more potent, in terms of life extension,than any of rapamycin, resveratrol, metformin, or spermidine. Referringto Table 11, it is clear that urolithins are the most potent among allof the compounds listed, in terms of life extension, increasing meanlife extension by 75% versus 27% for metformin, the most potent othercompound listed. This illustrates that urolithins show superior effectswith respect to their ability to delay the negative consequences linkedto aging. The effects of other urolithins have also shown impressiveeffects on life span extension (FIG. 4 ). Similar effects are seenacross all of the urolithins tested, so the compound class as a whole isexpected to show comparable effects.

TABLE 11 Comparison of the Effects of Compounds Active in Extending theLifespan of C. elegans Mean Life Extension Treatment C. elegans Dose (%)Reference Resveratrol Genotype: N2 100 μM 6.2-12.6 Wood J et al. (2004)Fed Live E. coli Nature 430:686-689. (OP50) Spermidine Genotype: N2 200μM 15 Eisenberg T et al.; Fed Live E. coli Nat. Cell Biol. (OP50)11:1305-1314. Rapamycin Genotype: N2 100 μM 19 Robida-Stubbs S et FedLive E. coli al. (2012) Cell (OP50) Metabolism 15:713-724. MetforminGenotype: N2  50 μM 27 Onken B et al. Fed Live E. coli (2010) PLoS ONE(OP50) 5(1):e8758. Urolithin A Genotype: N2  10 μM 27.7 This inventionFed Live E. coli  50 μM 75 (OP50)

As indicated above, the invention encompasses methods and compositionscombining urolithins, or precursors thereof, with one or more otheragents that promote autophagy. Such agents include, without limitation,rapamycin, metformin, resveratrol, spermidine, epigallocatechin gallate,genistein, silibinin, curcumin, clonidine, rilmenidine, tyramine,morphine, baclofen, mastoparan, propranolol, bupivacaine, N-dodecyllysinamide, tamoxifen, IFN-gamma, trehalose, and vinblastine.

The invention further encompasses methods and compositions combiningurolithins, or precursors thereof, with drugs reported to exertmitochondrial toxicity. Such toxic effects have been reported inconnection with, for example, valproic acid, antiretrovirals, statins,aspirin, aminoglycoside antibiotics, platinum-containingchemotherapeutic agents, acetaminophen, beta blockers, and steroids. Inthis setting, a urolithin or urolithin precursor is useful to counteractor treat the mitochondrial toxicity of the drugs reported to exertmitochondrial toxicity.

The invention further encompasses methods and compositions combiningurolithins, or precursors thereof, together with one or more agents thatare useful for mitochondrial biogenesis. Such compounds include, withoutlimitation, resveratrol, pyrroloquinoline quinone, genistein,hydroxyltyrosol, and quercetin.

The invention further encompasses methods and compositions combiningurolithins, or precursors thereof, together with one or more agents thatare useful for mitochondrial disorders. Such agents, which may beidentified as mitochondrial medications and supplements, includecoenzyme Q10 (CoQ10) as ubiquinol, CoQ10 as ubiquinone, riboflavin(vitamin B2), L-creatine, L-arginine, L-carnitine, B50 or B100 (Bvitamin complexes), vitamin E, vitamin C, alpha-lipoic acid, and folinicacid (e.g., as leucovorin).

The invention further encompasses methods and compositions combiningurolithins, or precursors thereof, together with one or more naturalproducts that are useful in the treatment of disorders of weightmanagement, metabolism, cognition, mood, and mitochondrial diseases.

In connection with disorders of weight management and metabolism, suchnatural products include, without limitation,23-oxo-3α-hydroxycycloart-24-en-26-oic acid (diabetes); berberine(diabetes, hypertension, and hypercholesterolemia); mangiferin(dyslipidemia); arabinoxylan (blood glucose management); beta-glucans(weight loss); chromium, fructose, hydroxypropylmethylcellulose,pectins, digestable starch, and konjacmannan (to promote weight loss);alpha-linolenic acid (ALA), beta-glucans, chitosan, guar gum, monacolinK, sterols, stanols, pectins, unsaturated fats, and linoleic acid (formanagement of blood cholesterol); calcium, chloride, lactulose, andfiber (for improved digestion); folate, choline, zinc, vitamin B6,magnesium, biotin, chromium, vitamin D (for improved metabolism);riboflavin (vitamin B2), vitamin A, copper, and vitamin D (for improvediron utilization); iron, and vitamin B6 (for oxygen transport); andcholine (for liver).

In connection with disorders of cognition and mood, such naturalproducts include, without limitation, extract of Bacopa monnieri,omega-3 fatty acids, flavonoids, vitamin D, and vitamin E(age-associated cognitive decline); pycnogenol (cognition duringstress); omega-3 fatty acids, and Lactobacillus rhamnosus bacteria (mooddisorders); biotin, folate, magnesium, niacin, thiamin, vitamin B6,vitamin B12, and vitamin C (psychological function); omega-3 fattyacids, curcumin, and vitamin E (traumatic brain injury); choline(seizure-induced memory impairment); omega-3 fatty acids, curcumin, andcopper (cognitive decay in Alzheimer's disease); danshensu (cognitivedefects secondary to diabetes); flavonoids, vitamin B6, vitamin B12,folate, caffeine, pantothenic acid, iodine, iron, and zinc (forcognitive enhancement); iron, and docosahexaenoic acid (DHA) (forcognitive development); and melatonin (for jet lag and for sleep).

In connection with mitochondrial diseases, such natural productsinclude, without limitation, ethanol extract of Cassia seed (liverprotection); viniferin (Huntington's disease); Korean mistletoe (Viscumalbum coloratura) extract (for increasing endurance capacity andreducing fatigue); β-phenylethyl isothiocyanate (PEITC), andepigallocatechin-3-gallate (EGCG) (for cancer); and coenzyme Q10,creatine, lipoic acid, and whey-based cysteine supplement (mitochondrialdiseases).

Formulations and Clinical Use

Methods and compositions of the invention are believed to be useful inany of a variety of clinical settings for which increasing autophagyand/or improving mitochondrial function are desirable. Methods andcompositions of the invention can be used for the treatment andprevention of diseases and conditions where increasing autophagy and/orimproving mitochondrial function are desirable. Non-limiting indicationsfor clinical use include: immune disorders, improving stem cellfunction, all aging-related health conditions where decline in autophagywith aging and/or increase in defective or suboptimally functioningmitochondria accumulate, e.g., neurodegenerative disease, heart disease,vascular disease, atherosclerosis, macular degeneration, sensory hearingloss, obesity, fatty liver, cancer, and infectious disease. Methods andcompositions of the invention are also believed to be useful in thetreatment and prevention of ischemia and reperfusion injury (including,for example, stroke, myocardial infarction, cardiac bypass, and organtransplantation). Methods and compositions of the invention are alsobelieved to be useful in the treatment of certain skin disorders,including skin aging, skin inflammation, and psoriasis. Methods andcompositions of the invention are also believed to be useful in thetreatment of osteoarthritis.

In another aspect, the present invention features a method of enhancingautophagy in a subject with a disease or condition; the method includesadministering to the subject an effective amount of a urolithin or aprecursor thereof, thereby treating the disease or condition in thesubject. In related embodiments, the invention includes methods oftreating or preventing any of a variety of diseases or conditions byproviding to a subject in need thereof an effective amount of aurolithin or precursor thereof. In particular embodiments, the urolithinor precursor thereof is present in a pharmaceutical composition. Incertain embodiments, the pharmaceutical composition comprises one ormore additional active agents, such as, e.g., one or more additionalurolithin or precursor thereof, or other compound described herein. Inparticular embodiments, the urolithin or precursor thereof is present ina medical food or beverage, functional food or beverage ornutraceutical. In certain embodiments, the medical food or beverage,functional food or beverage or nutraceutical comprises one or moreadditional active agents, such as, e.g., one or more additionalurolithin or precursor thereof, or other compound described herein.

In certain embodiments, the disease or condition is metabolic stress. Inparticular embodiments, the disease or disorder is nutrient deprivation,growth factor depletion, or hypoxia.

In certain embodiments, the disease or condition is a heart disease orcondition, such as, e.g., cardiac hypertrophy, left ventriculardilation, diminished cardiac output, decreased cardiac function, orDanon disease.

In certain embodiments, the disease or condition is muscular atrophy,e.g., muscle disuse atrophy, skeletal muscular atrophy, or sarcopenia ofaging.

In certain embodiments, the disease or condition is a muscledegenerative disorder, such as, e.g., a muscular dystrophy, e.g.,Duchenne's muscular dystrophy, or Ulrich myopathy.

In certain embodiments, the disease or condition is a liver disease,such as, e.g., cancer, cirrhosis, α1-antitrypsin deficiency,nonalcoholic fatty liver disease, alcoholic induced liver disease, ordrug induced liver injury.

In certain embodiments, the disease or condition is ischemic reperfusioninjury. In particular embodiments, the ischemic reperfusion injury isdue to liver surgery.

In certain embodiments, the disease or condition is a cardiac infarct.

In certain embodiments, the disease or condition relates to futuresurgical procedures and prevention or treatment of potential damage tocells and tissue, such as, e.g. angioplasty, heart valve repair, orcardiac bypass surgery.

In certain embodiments, the disease or condition is an inflammatorydisease or disorder. In particular embodiments, it is an autoimmunedisorder, such as, e.g., lupus erythematosus. In particular embodiments,it is an inflammatory disease or disorder, such as, e.g. psoriasis.

In certain embodiments, the disease or condition is a neurodegenerativedisease or disorder, such as, e.g., Parkinson's, Alzheimer's,Huntington's or polyQ disease.

In certain embodiments, the disease or condition is a cancer or tumor.

In one embodiment of this aspect, the disease is caused by misfoldedprotein aggregates. In another embodiment of this aspect, the diseasecaused by misfolded protein aggregates is selected from the groupincluding: Alzheimer's disease, Parkinson's disease, amyotrophic lateralsclerosis, Huntington's disease, spinocerebellar ataxia, oculopharyngealmuscular dystrophy, prion diseases, fatal familial insomnia, alpha-1antitrypsin deficiency, dentatorubral pallidoluysian atrophy, frontaltemporal dementia, progressive supranuclear palsy, X-linked spinobulbarmuscular atrophy, and neuronal intranuclear hyaline inclusion disease.In a further embodiment of this aspect, the disease associated withmisfolded protein aggregates is a chronic disease. In yet anotherembodiment of this aspect, the disease is cancer.

In addition to their usefulness in extending longevity, methods andcompositions of the invention are also believed to be useful forextending health span. As used herein, “health span” (or, equivalently,“healthspan”) refers to the period of an individual's life during whichthey are generally healthy and free from serious or chronic illness. Inone embodiment, “health span” (or, equivalently, “healthspan”) refers tothe period of time during which an individual meets one or more selectedmeasures of health. An increase in health span refers to an extension inthe period of health, according to such measures, as compared to theperiod of health in a control population. An increase in health span canbe measured, e.g., by determining the length of time that an individualcontinues to meet the selected measure(s) of health.

The urolithin or precursor thereof may be administered, alone ortogether with at least one other agent, to a subject (e.g., a mammal) inany of a variety of ways. For example, the urolithin or precursorthereof can be administered orally or parenterally. Parenterallyincludes, without limitation, intravenously, intramuscularly,intraperitoneally, subcutaneously, intra-articularly, intrasynovially,intraocularly, intrathecally, topically, or by inhalation. As such, theform of the urolithin or precursor thereof dose can be any of a varietyof suitable forms, including neat compounds, natural foods, processedfoods, natural juices, concentrates and extracts, injectable solutions,microcapsules, nano-capsules, liposomes, plasters, inhalation forms,nose sprays, nosedrops, eyedrops, sublingual tablets, andsustained-release preparations.

The compounds of this invention can be provided in isolated form. Asused herein, the term “isolated” means substantially removed from othercompounds or components with which the compound of interest mayotherwise be found, for example, as found in nature. In one embodiment,a compound is isolated when it is essentially completely removed fromother compounds or components with which the compound of interest mayotherwise be found. In one embodiment, a compound is isolated when it ispure.

The compounds of this invention can be incorporated into a variety offormulations for therapeutic administration. More particularly, thecompounds of the present invention can be formulated into pharmaceuticalcompositions by combination with appropriate pharmaceutically acceptablecarriers or diluents, and may be formulated into preparations in solid,semi-solid, liquid or gaseous forms, such as tablets, capsules, powders,granules, ointments, solutions, suppositories, injections, inhalants,gels, microspheres, and aerosols. As such, administration of thecompounds can be achieved in various ways, including without limitationoral, buccal, rectal, intravenous, intramuscular, intraperitoneal,intradermal, transdermal, and intratracheal administration. The activeagent may be systemic after administration or may be localized by theuse of regional administration, intramural administration, or use of animplant that acts to retain the active dose at the site of implantation.

The compounds of the invention can also be formulated as food additives,food ingredients, functional foods, dietary supplements, medical foods,nutraceuticals, or food supplements. In certain embodiments, compoundsof the invention can be included into nutraceutical beverages of varyingvolumes to permit the administration of a daily dose in a convenientformat. As a non-limiting example, beverages may deliver effective dosesin a final volume ranging from 5 mL to 1,000 mL, delivered as a singledose or multiple doses. In certain embodiments, compositions and methodsof the invention are utilized for and in non-human animals. Accordingly,compounds and compositions of the invention may be formulated asveterinary products. Compounds and composition may also be formulatedinto functional foods for administration to animals, for example, dogs,cats, horses, etc.

In pharmaceutical dosage forms, the compounds may be administered in theform of their pharmaceutically acceptable salts. The term“pharmaceutically acceptable salt” is intended to encompass any and allacceptable salt forms derived from a physiologically acceptable acid orbase. The compounds of the present invention may be utilized as the freeacid or free base. Alternatively, the compounds of this invention may beused in the form of acid or base addition salts, which may be formed bymethods well known in the art.

The compounds may also be used in appropriate association with otherpharmaceutically active compounds. The following methods and excipientsare merely exemplary and are in no way limiting.

For oral preparations, the compounds can be used alone or in combinationwith appropriate additives to make tablets, powders, granules orcapsules, for example, with conventional additives, such as lactose,mannitol, corn starch or potato starch; with binders, such ascrystalline cellulose, cellulose derivatives, acacia, corn starch orgelatins; with disintegrators, such as corn starch, potato starch orsodium carboxymethylcellulose; with lubricants, such as talc ormagnesium stearate; and if desired, with diluents, buffering agents,moistening agents, preservatives and flavoring agents.

The compounds can be formulated into preparations for injections bydissolving, suspending or emulsifying them in an aqueous or nonaqueoussolvent, such as vegetable or other similar oils, synthetic aliphaticacid glycerides, esters of higher aliphatic acids or propylene glycol;and if desired, with conventional, additives such as solubilizers,isotonic agents, suspending agents, emulsifying agents, stabilizers andpreservatives.

The compounds can be utilized in aerosol formulation to be administeredvia inhalation. The compounds of the present invention can be formulatedinto pressurized acceptable propellants such as dichlorodifluoromethane,propane, nitrogen and the like.

Furthermore, the compounds can be made into suppositories by mixing witha variety of bases such as emulsifying bases or water-soluble bases. Thecompounds of the present invention can be administered rectally via asuppository. The suppository can include vehicles such as cocoa butter,carbowaxes and polyethylene glycols, which melt at body temperature, yetare solidified at room temperature.

Unit dosage forms for oral or rectal administration such as syrups,elixirs, and suspensions may be provided wherein each dosage unit, forexample, teaspoonful, tablespoonful, tablet or suppository, contains apredetermined amount of the composition containing one or more compoundsof the present invention. Similarly, unit dosage forms for injection orintravenous administration may comprise the compound of the presentinvention in a composition as a solution in sterile water, normal salineor another pharmaceutically acceptable carrier, wherein each dosageunit, for example, mL or L, contains a predetermined amount of thecomposition containing one or more compounds of the present invention.

Implants for sustained release formulations are well-known in the art.Implants are formulated as microspheres; slabs, etc., with biodegradableor non-biodegradable polymers. For example, polymers of lactic acidand/or glycolic acid form an erodible polymer that is well-tolerated bythe host. The implant containing the inhibitory compounds may be placedin proximity to a site of interest, so that the local concentration ofactive agent is increased relative to the rest of the body.

The term “unit dosage form”, as used herein, refers to physicallydiscrete units suitable as unitary dosages for human and animalsubjects, each unit containing a predetermined quantity of compounds ofthe present invention calculated in an amount sufficient to produce thedesired effect in association with a pharmaceutically acceptablediluent, carrier or vehicle. The specifications for the novel unitdosage forms of the present invention depend on the particular compoundemployed and the effect to be achieved, and the pharmacodynamicsassociated with each compound in the host.

The pharmaceutically acceptable excipients, such as vehicles, adjuvants,carriers or diluents, are readily available to the public. Moreover,pharmaceutically acceptable auxiliary substances, such as pH adjustingand buffering agents, tonicity adjusting agents, stabilizers, wettingagents and the like, are readily available to the public.

For clinical use, the urolithin or urolithin precursor is administeredin a therapeutically effective amount. As used herein, an “effectiveamount” refers to an amount that is sufficient to achieve or realize aspecified or desired biological effect. As used herein, a“therapeutically effective amount” refers to an amount sufficient torealize, in a single dose or multiple doses, a desired therapeuticeffect. A skilled artisan can determine therapeutically effectiveamounts based on in vitro, preclinical, or clinical studies, or anycombination thereof.

In one embodiment urolithins and their precursors can be delivered bymeans of natural products. One of these is the pomegranate, whichcontains ellagitannins and ellagic acid. Pomegranates may be processedin several ways for practicing this invention. Pomegranate juice canvary widely in terms of its polyphenol constituents, particularlypunicalagin, the ellagitannin having the highest concentration found inthe pomegranate. The starting material (i.e., pomegranates and theirvariety), the method of juicing, as well as the storage conditions willaffect punicalagin levels found in the final juice product. Table 12below provides examples of juices that have been store bought and ofdifferent origins. A large variation in punicalagin from one juice batchto another is observed. For the purpose of practicing this invention itis advantageous that individuals consume a standardized batch of juicethat is delivering known and consistent levels of ellagitannins.Presently in the marketplace pomegranate juice is not standardized byellagitannin levels and dosing is not indicated. Consequently, currentlyavailable juices are not suitable for effectively practicing theinvention as they do not offer the possibility of consistent dosing byan individual, due to the wide variations of ellagitannins and theirconcentrations found in the different products.

TABLE 12 Variation of Punicalagin Content in Different CommerciallyAvailable Pomegranate Juices Commercial Punicalagins Juice (g/L) 1 0.082 0.28 3 0.04 4 0.32 5 0.26 6 0.89 7 0.10 8 0.10 9 0.02 10 0.51 11 0.38

As described earlier, the consumption of ellagitannins by humans leadsto the formation of urolithins by the gut microflora prior to theirabsorption. It is believed that consumption of ellagitannins holds thepotential to modify the microflora of the gut. In one study in rats itwas observed that urolithins were found in the urine only following fourdays of consumption, suggesting that the rats required time to adapttheir microflora to metabolize pomegranate tannins to produce theurolithin metabolites. Cerda et al. (2003) Eur J Nutr 42:18-28. Thisfinding suggests that optimal dosing of ellagitannins in humans mayrequire a similar period for each individual's microflora to adaptoptimally to metabolize consumed ellagitanins or ellagic acidequivalents into urolithins.

With urolithin precursors, including but not limited to ellagitanninsand ellagic acid, a minimal treatment period sometimes may be neededprior to observing autophagy or the health benefits, depending on thesubject. For example, this pre-treatment period may be 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, or 31 days, or 1, 2, or 3 months.

Additionally, it has been shown that individuals' microfloracompositions can vary dramatically and that some individuals maymetabolize ellagitannins more completely than others. For instance,following the consumption of pomegranate juice, some individuals maypreferentially convert ellagitannins or ellagic acid equivalents intourolithin A, while others may preferentially convert into urolithin C.Also, it has been observed that certain individuals are high converters(i.e., a high level of urolithins are formed in the gut from consumedellagitannins and ellagic acid equivalents), while others are lowconverters (i.e., only a low level of urolithins are formed in the gutfrom consumed ellagitannins and ellagic acid equivalents). Thesedifferences in metabolite production from high producer to low producerand variations of urolithin are believed to be due to differences incolonic microflora and the potential requirement for an adaptationperiod. Cerda et al. (2004) Eur J Nutr 43:205-220. Consequently, due tothese variations when administering ellagitannins and their equivalents,for certain applications it may be beneficial to administer urolithinsdirectly.

Dosing will generally be daily to weekly. In one embodiment, dosing isat least weekly. For example, a subject may receive one dose onceweekly, twice weekly, thrice weekly, or every other day. In oneembodiment, dosing is at least daily. For example, a subject may receiveone or more doses daily.

It is believed that dosing for greatest efficacy in humans involvesextended, daily administration, e.g., when delivering ellagitannins,amounts equivalent to at least 16 ounces (500 mL) of pomegranate juiceper day. Extended use is contemplated to include use for 1 month, 2months, 3 months, 4 months, 5 months, 6 months, or even longer.

For clinical use, a urolithin or precursor may be administered to treateither a chronic or an acute condition. As autophagy is observed to beincreased in cells in less than one day, for example after 8 hours,urolithins and their variants of Formula I, Formula II and Formula IIImay be administered in an acute fashion to treat a condition acutely inneed an induction of autophagy. Such instances may include conditionsdue to reperfusion injuries (for example, organ transplantation), heartattack, stroke, ischemic insult, during surgery (for example, during anangioplasty procedure or replacement of a heart valve), or following atraumatic injury.

For clinical use, a urolithin or precursor thereof will generally beadministered in an amount equal or equivalent to 0.2-2000 milligram (mg)of urolithin per kilogram (kg) of body weight of the subject per day. Inone embodiment, the urolithin or precursor thereof is administered in adose equal or equivalent to 2-2000 mg of urolithin per kg body weight ofthe subject per day. In one embodiment, the urolithin or precursorthereof is administered in a dose equal or equivalent to 20-2000 mg ofurolithin per kg body weight of the subject per day. In one embodiment,the urolithin or precursor thereof is administered in a dose equal orequivalent to 50-2000 mg of urolithin per kg body weight of the subjectper day. In one embodiment, the urolithin or precursor thereof isadministered in a dose equal or equivalent to 100-2000 mg of urolithinper kg body weight of the subject per day. In one embodiment, theurolithin or precursor thereof is administered in a dose equal orequivalent to 200-2000 mg of urolithin per kg body weight of the subjectper day. Where a precursor of urolithin is to be administered ratherthan a urolithin, it is administered in an amount that is equivalent to,i.e., sufficient to deliver, the above-stated amounts of urolithin.

The formulations of urolithin or precursor thereof can be administeredto human subjects in therapeutically effective amounts. Typical doseranges are from about 0.01 microgram/kg to about 2 mg/kg of body weightper day. The dosage of drug to be administered is likely to depend onsuch variables as the type and extent of the disorder, the overallhealth status of the particular subject, the specific compound beingadministered, the excipients used to formulate the compound, and itsroute of administration. Routine experiments may be used to optimize thedose and dosing frequency for any particular compound.

In one embodiment, the urolithin or precursor thereof is administered ata concentration in the range from about 0.001 microgram/kg to greaterthan about 500 mg/kg. For example, the concentration may be 0.001microgram/kg, 0.01 microgram/kg, 0.05 microgram/kg, 0.1 microgram/kg,0.5 microgram/kg, 1.0 microgram/kg, 10.0 microgram/kg, 50.0microgram/kg, 100.0 microgram/kg, 500 microgram/kg, 1.0 mg/kg, 5.0mg/kg, 10.0 mg/kg, 15.0 mg/kg, 20.0 mg/kg, 25.0 mg/kg, 30.0 mg/kg, 35.0mg/kg, 40.0 mg/kg, 45.0 mg/kg, 50.0 mg/kg, 60.0 mg/kg, 70.0 mg/kg, 80.0mg/kg, 90.0 mg/kg, 100.0 mg/kg, 150.0 mg/kg, 200.0 mg/kg, 250.0 mg/kg,300.0 mg/kg, 350.0 mg/kg, 400.0 mg/kg, 450.0 mg/kg, to greater thanabout 500.0 mg/kg or any incremental value thereof. It is to beunderstood that all values and ranges between these values and rangesare meant to be encompassed by the present invention.

In one embodiment, the urolithin or precursor thereof is administered ata dosage in the range from about 0.2 milligram/kg/day to greater thanabout 100 mg/kg/day. For example, the dosage may be 0.2 mg/kg/day to 100mg/kg/day, 0.2 mg/kg/day to 50 mg/kg/day, 0.2 mg/kg/day to 25 mg/kg/day,0.2 mg/kg/day to 10 mg/kg/day, 0.2 mg/kg/day to 7.5 mg/kg/day, 0.2mg/kg/day to 5 mg/kg/day, 0.25 mg/kg/day to 100 mg/kg/day, 0.25mg/kg/day to 50 mg/kg/day, 0.25 mg/kg/day to 25 mg/kg/day, 0.25mg/kg/day to 10 mg/kg/day, 0.25 mg/kg/day to 7.5 mg/kg/day, 0.25mg/kg/day to 5 mg/kg/day, 0.5 mg/kg/day to 50 mg/kg/day, 0.5 mg/kg/dayto 25 mg/kg/day, 0.5 mg/kg/day to 20 mg/kg/day, 0.5 mg/kg/day to 15mg/kg/day, 0.5 mg/kg/day to 10 mg/kg/day, 0.5 mg/kg/day to 7.5mg/kg/day, 0.5 mg/kg/day to 5 mg/kg/day, 0.75 mg/kg/day to 50 mg/kg/day,0.75 mg/kg/day to 25 mg/kg/day, 0.75 mg/kg/day to 20 mg/kg/day, 0.75mg/kg/day to 15 mg/kg/day, 0.75 mg/kg/day to 10 mg/kg/day, 0.75mg/kg/day to 7.5 mg/kg/day, 0.75 mg/kg/day to 5 mg/kg/day, 1.0 mg/kg/dayto 50 mg/kg/day, 1.0 mg/kg/day to 25 mg/kg/day, 1.0 mg/kg/day to 20mg/kg/day, 1.0 mg/kg/day to 15 mg/kg/day, 1.0 mg/kg/day to 10 mg/kg/day,1.0 mg/kg/day to 7.5 mg/kg/day, 1.0 mg/kg/day to 5 mg/kg/day, 2mg/kg/day to 50 mg/kg/day, 2 mg/kg/day to 25 mg/kg/day, 2 mg/kg/day to20 mg/kg/day, 2 mg/kg/day to 15 mg/kg/day, 2 mg/kg/day to 10 mg/kg/day,2 mg/kg/day to 7.5 mg/kg/day, or 2 mg/kg/day to 5 mg/kg/day.

In one embodiment, the urolithin or precursor thereof is administered ata dosage in the range from about 0.25 milligram/kg/day to about 25mg/kg/day. For example, the dosage may be 0.25 mg/kg/day, 0.5 mg/kg/day,0.75 mg/kg/day, 1.0 mg/kg/day, 1.25 mg/kg/day, 1.5 mg/kg/day, 1.75mg/kg/day, 2.0 mg/kg/day, 2.25 mg/kg/day, 2.5 mg/kg/day, 2.75 mg/kg/day,3.0 mg/kg/day, 3.25 mg/kg/day, 3.5 mg/kg/day, 3.75 mg/kg/day, 4.0mg/kg/day, 4.25 mg/kg/day, 4.5 mg/kg/day, 4.75 mg/kg/day, 5 mg/kg/day,5.5 mg/kg/day, 6.0 mg/kg/day, 6.5 mg/kg/day, 7.0 mg/kg/day, 7.5mg/kg/day, 8.0 mg/kg/day, 8.5 mg/kg/day, 9.0 mg/kg/day, 9.5 mg/kg/day,10 mg/kg/day, 11 mg/kg/day, 12 mg/kg/day, 13 mg/kg/day, 14 mg/kg/day, 15mg/kg/day, 16 mg/kg/day, 17 mg/kg/day, 18 mg/kg/day, 19 mg/kg/day, 20mg/kg/day, 21 mg/kg/day, 22 mg/kg/day, 23 mg/kg/day, 24 mg/kg/day, 25mg/kg/day, 26 mg/kg/day, 27 mg/kg/day, 28 mg/kg/day, 29 mg/kg/day, 30mg/kg/day, 31 mg/kg/day, 32 mg/kg/day, 33 mg/kg/day, 34 mg/kg/day, 35mg/kg/day, 36 mg/kg/day, 37 mg/kg/day, 38 mg/kg/day, 39 mg/kg/day, 40mg/kg/day, 41 mg/kg/day, 42 mg/kg/day, 43 mg/kg/day, 44 mg/kg/day, 45mg/kg/day, 46 mg/kg/day, 47 mg/kg/day, 48 mg/kg/day, 49 mg/kg/day, or 50mg/kg/day.

In another embodiment, the urolithin or precursor thereof isadministered in concentrations that range from 0.01 micromolar togreater than or equal to 500 micromolar. For example, the dose may be0.01 micromolar, 0.02 micromolar, 0.05 micromolar, 0.1 micromolar, 0.15micromolar, 0.2 micromolar, 0.5 micromolar, 0.7 micromolar, 1.0micromolar, 3.0 micromolar, 5.0 micromolar, 7.0 micromolar, 10.0micromolar, 15.0 micromolar, 20.0 micromolar, 25.0 micromolar, 30.0micromolar, 35.0 micromolar, 40.0 micromolar, 45.0 micromolar, 50.0micromolar, 60.0 micromolar, 70.0 micromolar, 80.0 micromolar, 90.0micromolar, 100.0 micromolar, 150.0 micromolar, 200.0 micromolar, 250.0micromolar, 300.0 micromolar, 350.0 micromolar, 400.0 micromolar, 450.0micromolar, to greater than about 500.0 micromolar or any incrementalvalue thereof. It is to be understood that all values and ranges betweenthese values and ranges are meant to be encompassed by the presentinvention.

In yet another embodiment, the urolithin or precursor thereof isadministered at concentrations that range from 0.10 microgram/mL to500.0 microgram/mL. For example, the concentration may be 0.10microgram/mL, 0.50 microgram/mL, 1 microgram/mL, 2.0 microgram/mL, 5.0microgram/mL, 10.0 microgram/mL, 20 microgram/mL, 25 microgram/mL. 30microgram/mL, 35 microgram/mL, 40 microgram/mL, 45 microgram/mL, 50microgram/mL, 60.0 microgram/mL, 70.0 microgram/mL, 80.0 microgram/mL,90.0 microgram/mL, 100.0 microgram/mL, 150.0 microgram/mL, 200.0microgram/mL, 250.0 g/mL. 250.0 microgram/mL, 300.0 microgram/mL, 350.0microgram/mL, 400.0 microgram/mL, 450.0 microgram/mL, to greater thanabout 500.0 microgram/mL or any incremental value thereof. It is to beunderstood that all values and ranges between these values and rangesare meant to be encompassed by the present invention.

The term “effective amount” refers to the amount of the urolithin orprecursor thereof required to enhance autophagy, e.g., in a diseaseassociated with misfolded protein aggregates. The effective amount ofthe urolithin or precursor thereof used to enhance autophagy variesdepending upon the manner of administration, the age, body weight, andgeneral health of the subject. An effective amount of the urolithin orprecursor thereof, as defined herein, may vary according to factors suchas the disease state, age, and weight of the subject, and the ability ofthe urolithin or precursor thereof to elicit a desired response in thesubject. Dosage regimens may be adjusted to provide the optimumtherapeutic response. An effective amount is also one in which any toxicor detrimental effects (e.g., side effects) of the autophagy inducingcompound are outweighed by the therapeutically beneficial effects. Forexample, a therapeutically effective amount of the urolithin orprecursor thereof (i.e., an effective dosage) may range from about 0.001to 30 mg/kg body weight, preferably about 0.01 to 25 mg/kg body weight,more preferably about 0.1 to 20 mg/kg body weight, and even morepreferably about 1 to 10 mg/kg, 2 to 9 mg/kg, 3 to 8 mg/kg, 4 to 7mg/kg, or 5 to 6 mg/kg body weight. The skilled artisan will appreciatethat certain factors may influence the dosage required to effectivelytreat a subject, including but not limited to the severity of thedisease or disorder, previous treatments, the general health and/or ageof the subject, and other diseases present. Moreover, treatment of asubject with a therapeutically effective amount of the urolithin orprecursor thereof can include a single treatment or, preferably, caninclude a series of treatments. In one example, a subject is treatedwith the urolithin or precursor thereof in the range of between about0.1 to 20 mg/kg body weight, one time per week for between about 1 to 10weeks, preferably between 2 to 8 weeks, more preferably between about 3to 7 weeks, and even more preferably for about 4, 5, or 6 weeks. It willalso be appreciated that the effective dosage of the urolithin orprecursor thereof used for treatment may increase or decrease over thecourse of a particular treatment.

Non-limiting, illustrative examples of ellagitannin, ellagic acid, andurolithin dosages that may be used are provided in Table 13.

TABLE 13 Ellagitannin/Ellagic Acid/Urolithin Dosing in Mice and Man MiceMan Punicalagin (mg/kg/d) 90.00 6.56 Punicalagin (moles/kg/day) 97.657.11 Ellagic acid (mg/kg/day) 75.00 5.46 Ellagic acid (moles/kg/day)22.65 1.65 Total Urolithins (mg/kg/day) 55.00 4.01 Urolithin A(moles/kg/day) 12.49 0.91 Urolithin B (moles/kg/day) 11.66 0.85Urolithin C (moles/kg/day) 13.42 0.98 Urolithin D (moles/kg/day) 14.251.04

Additional non-limiting, illustrative examples of urolithin doses thatmay be used are provided in Table 14.

TABLE 14 Urolithin Dosing in Mouse and Human Mouse Dose Human Dose(mg/kg/day) (mg/kg/day) 5 0.36 10 0.73 15 1.09 20 1.46 30 2.19 40 2.9150 3.64 60 4.37 70 5.10 80 5.33 90 6.56 100 7.29 110 8.01 120 8.74 1309.47 140 10.20 150 10.93 160 11.66 170 12.38 180 13.11 190 13.84 20014.57 210 15.30 220 16.03 230 16.76 240 17.48 250 18.21 260 18.94 27019.67 280 20.40 290 21.13 300 21.86 310 22.58 320 23.31 330 24.04 34024.77 350 25.50 360 26.23 370 26.96 380 27.68 390 28.41 400 29.14 41029.87 420 30.60 430 31.33

Additional non-limiting, illustrative examples of urolithin dose rangesthat may be used are provided in Table 15.

TABLE 15 Urolithin Dosing in Humans Human Dose Ranges (mg/kg/day) LowHigh 0.2 4 0.2 5 0.2 6 0.2 7 0.2 8 0.2 9 0.2 10 0.2 15 0.2 20 0.2 25 0.230 0.2 35 0.2 40 0.5 2 0.5 3 0.5 4 0.5 5 0.5 10 0.5 15 0.5 20 0.5 25 0.530 0.5 35 0.5 40 0.75 5 0.75 10 0.75 15 0.75 20 0.75 25 0.75 30 0.75 350.75 40 1 2 1 3 1 4 1 5 1 6 1 7 1 8 1 9 1 10 1 11 1 12 1.25 4 1.25 5

Any dose may be given as a single dose or as divided doses.

In one embodiment, the urolithin or precursor thereof is administered ina dose sufficient to achieve a peak serum level of urolithin and itsknown metabolites (glucoronides, sulfonates, etc.) of at least 0.001micromolar (μM). In one embodiment, the urolithin or precursor thereofis administered in a dose sufficient to achieve a peak serum level ofurolithin of at least 0.01 μM. In one embodiment, the urolithin orprecursor thereof is administered in a dose sufficient to achieve a peakserum level of urolithin of at least 0.1 μM. In one embodiment, theurolithin or precursor thereof is administered in a dose sufficient toachieve a peak serum level of urolithin of at least 1 μM. In variousembodiments, the urolithin or precursor thereof is administered in adose sufficient to achieve a peak serum level of urolithin of at least10 μM, at least 20 μM, at least 30 μM, at least 40 μM, at least 50 μM,at least 60 μM, at least 70 μM, at least 80 μM, at least 90 μM, at least100 μM, or at least 200 μM.

In one embodiment, the urolithin or precursor thereof is administered ina dose sufficient to achieve a sustained serum level of urolithin of atleast 0.001 micromolar (μM). In one embodiment, the urolithin orprecursor thereof is administered in a dose sufficient to achieve asustained serum level of urolithin of at least 0.01 μM. In oneembodiment, the urolithin or precursor thereof is administered in a dosesufficient to achieve a sustained serum level of urolithin of at least0.1 μM. In one embodiment, the urolithin or precursor thereof isadministered in a dose sufficient to achieve a sustained serum level ofurolithin of at least 1 μM. In one embodiment, the urolithin orprecursor thereof is administered in a dose sufficient to achieve asustained serum level of urolithin of at least 10 μM, the urolithin orprecursor thereof is administered in a dose sufficient to achieve asustained serum level of urolithin of at least 50 μM. The sustainedserum level can be measured using any suitable method, for example, highpressure liquid chromatography (HPLC) or HPLC-MS.

EXAMPLES Example 1 Urolithin A Synthesis

Urolithin A (4) was prepared in two steps starting from bromide 1 andresorcinol 2. The pure compound was obtained as a pale yellow powder.

Step 1:

A mixture of 2-bromo-5-methoxybenzoic acid 1 (27.6 g; 119 mmol; 1.0eq.), resorcinol 2 (26.3 g; 239 mmol; 2.0 eq.) and sodium hydroxide(10.5 g; 263 mmol; 2.2 eq.) in water (120 mL) was heated under refluxfor 1 hour. A 5% aqueous solution of copper sulphate (3.88 g ofCuSO₄·5H₂O in 50 mL water; 15.5 mmol; 0.1 eq.) was then added and themixture was refluxed for additional 30 minutes. The mixture was allowedto cool to room temperature and the solid was filtered on a Büchnerfilter. The residue was washed with cold water to give a pale red solidwhich was triturated in hot MeOH. The suspension was left overnight at4° C. The resultant precipitate was filtered and washed with cold MeOHto yield the title compound 3 as a pale brown solid.

Step 2:

To a suspension of 3 (10.0 g; 41 mmol; 1.0 eq.) in dry dichloromethane(100 mL) was added dropwise at 0° C. a 1 M solution of boron tribromidein dry dichloromethane (11.93 mL of pure BBr₃ in 110 mL of anhydrousdichloromethane; 124 mmol; 3.0 eq.). The mixture was left at 0° C. for 1hour and was then allowed to warm up to room temperature. The solutionwas stirred at that temperature for 17 hours. Then ice was addedthoroughly to the mixture. The yellow precipitate was filtered andwashed with cold water to give a yellow solid which was heated to refluxin acetic acid for 3 hours. The hot solution was filtered quickly andthe precipitate was washed with acetic acid, then with diethyl ether toyield the title compound 4 as a yellow solid. ¹H and ¹³C NMR were inaccordance with the structure of 4.

Example 2 Urolithin B Synthesis

Urolithin B (3) was prepared in one step by coupling of resorcinol (1)and 2-bromobenzoic acid (2) following the procedure employed for thepreparation of urolithin A in Example 1.

A mixture of resorcinol 1 (8.40 g; 75.6 mmol; 2.0 eq.), 2-bromobenzoicacid (7.60 g; 37.8 mmol; 1.0 eq.) and NaOH (3.34 g; 83.5 mmol; 2.2 eq.)in water (38 mL) was heated at reflux and stirred for 2 hours. Water (30mL) and CuSO₄·5H₂O (0.95 g; 3.78 mmol; 0.1 eq.) were added and themixture was stirred under reflux for an additional 1 hour. The reactionwas then cooled to room temperature and the precipitate was filtered.The product was then dissolved in absolute ethanol and concentrated. Thecrude was dissolved in hot methanol and filtered on paper to afford thetitle compound. ¹H and ¹³C NMR were in accordance with the structure of3.

Example 3 Urolithin C Synthesis

Urolithin C (4) was prepared in two steps starting from resorcinol (1)and 2-bromo-4,5-dimethoxybenzoic acid (2) following the procedure abovefor the preparation of urolithin A in Example 1.

Step 1: Preparation of Compound 3

A mixture of resorcinol 1 (8.00 g; 72.6 mmol; 2.0 eq.),2-bromo-4,5-dimethoxybenzoic acid 2 (9.50 g; 36.3 mmol; 1.0 eq.) andNaOH (3.20 g; 79.9 mmol; 2.2 eq.) in water (36 mL) was heated at refluxand stirred for 2 hours. Water (20 mL) and CuSO₄·5H₂O (0.91 g; 3.63mmol; 0.1 eq.) were added and the mixture was stirred under reflux foran additional 1 hour. The reaction was then cooled down to roomtemperature and the precipitate was filtered. The product was thendissolved in absolute ethanol and concentrated. The crude was engaged inthe next step without further purification.

Step 2: Preparation of Compound 4

A 1M solution of BBr₃ in dichloromethane (11 mL; 11.0 mmol; 6.0 eq.) wasadded dropwise to a solution of intermediate 3 (500 mg; 1.84 mmol; 1.0eq.) in cold (0° C.) dichloromethane (5 mL). The reaction mixture wasstirred at 0° C. during 1 hour and at room temperature for an additional48 hours. The reaction was then hydrolysed by addition of ice. Theprecipitate was filtered and washed with ice-cold water until pH 7. Theproduct was then dissolved in absolute ethanol and concentrated. Thecrude was dried under vacuum in presence of P₂O₅ to afford the titlecompound. ¹H and ¹³C NMR were in accordance with the structure of 4.

Example 4 Urolithin D Synthesis

Urolithin D (5) was prepared in three steps starting from2,3-dimethoxyphenol (1) and 2-bromo-4,5-dimethoxybenzoic acid (2).

Step 1: Preparation of Compound 3

Oxalyl chloride (1.07 mL; 12.6 mmol; 1.1 eq.) and dimethylformamide (1drop; cat.) were added to a cold (0° C.) suspension of2-bromo-4,5-dimethoxybenzoic acid 2 (3.00 g; 11.5 mmol; 1.0 eq.) indichloromethane (20 mL). After addition, the reaction mixture wasstirred for 5 minutes at 0° C. and then for 16 hours at roomtemperature. The reaction mixture was concentrated under reducedpressure. The resulting oil was solubilized in dichloromethane (10 mL)and cooled to 0° C. 2,3-Methoxyphenol 1 (1.90 g; 12.6 mmol; 1.1 eq.) andtriethylamine (2.4 mL; 17.2 mmol; 1.5 eq.) were successively added andthe mixture was stirred for 5 minutes. Acetonitrile (10 mL) was finallyadded and the resulting solution was stirred for 16 hours at roomtemperature. The reaction was quenched by addition of a saturatedsolution of ammonium chloride and the layers were separated. The aqueousphase was extracted with dichloromethane and the combined organic layerswere dried over sodium sulphate, filtered and concentrated under reducedpressure. The crude was purified by column chromatography on silica gelto yield the title compound 3.

Step 2: Preparation of Compound 4

Sodium acetate (1.79 g; 21.8 mmol; 2.0 eq.), (S)-Phos (451 mg; 1.1 mmol;0.1 eq.) and palladium diacetate (244 mg; 1.1 mmol; 0.1 eq.) weresuccessively added to a solution of 3 (4.32 g; 10.9 mmol; 1 eq.) indimethylacetamide (200 mL) under an argon atmosphere. The reactionmixture was stirred at 130° C. for 3 days. The reaction was quenched byaddition of water and the mixture was extracted with dichloromethane.The combined organic layers were washed with brine, dried over sodiumsulphate, and concentrated under reduced pressure. The remainingdimethylacetamide was eliminated using a Hickman apparatus. The crudedark oil was purified by column chromatography on silica gel to yieldthe title compound 4.

Step 3: Preparation of Compound 5

Boron tribromide (726 μL; 10.3 mmol; 5.0 eq.) was added to a cold (15°C.) solution of compound 4 (650 mg; 2.0 mmol; 1.0 eq.) in chloroform(7.3 mL). After the addition, the reaction mixture was allowed to reachroom temperature and stirred for 15 minutes. The mixture was then heatedup to 60° C. and stirred for 4 days. The reaction was quenched byaddition of methanol and the mixture was evaporated to dryness. Thecrude was purified by column chromatography on C18 to yield purecompound 5.

Example 5

Urolithins Extend Lifespan in C. elegans

C. elegans strains were cultured at 20° C. on nematode growth media(NGM) agar plates seeded with E. coli strain OP50. Strain used waswild-type Bristol N2 provided by the Caenorhabditis Genetics Center(University of Minnesota). All Compounds were dissolved in DMSO. Animalswere exposed to compounds from eggs on plates seeded with live OP50bacteria. Control plates were prepared with the correspondingconcentrations of DMSO (0.1%). For lifespan tests, worms were observedusing a Nikon SMZ1500 stereomicroscope (Nikon, Melville, N.Y., USA).Briefly, 10 L4 worms were transferred on plates containing NGM mediumwith either vehicle alone (DMSO 0.1%) or test compounds dissolved inDMSO (DMSO 0.1%) and seeded with E. coli strain OP50. After 3 days(corresponding to day 0), newly developed 60 to 100 L4 worms were usedper condition, scored and transferred to fresh plates every 3 daysstarting from day 1 (young adult worms). All lifespan experiments wereperformed at 20° C. Animals that crawled off the plate or had an“exploded vulva” phenotype were censored.

Treatment with urolithins A, B, C, and D at a concentration of 50 μMextended the lifespan in C. elegans (FIG. 4 ). Ellagic acid (EA) at thesame concentration showed no effects in prolonging the lifespan of theC. elegans.

Treatment with urolithin A extended the lifespan in C. elegans in adose-dependent manner (FIG. 5 ). At a concentration of 10 μM the medianlifespan increased by 27.7%, and at a concentration of 50 μM the medianlifespan increased by 75%.

Example 6 Urolithin A Activates AMPK/aak-2 and SIRT1/Sir-2.1 to MediateLongevity

Lifespan assays were performed with (A) wild-type (N2), (B)sir-2.1(ok434) mutant, (C) aak-2(ok524), (D) daf-16(mu86), (E)eat-2(ad465) and (F) daf-2(e1370) mutant strains of C. elegansmaintained during their full life on plates supplemented with either 50μM of urolithin A or with a control treatment and seeded with OP50bacteria. Control plates without urolithin A contained an equivalentconcentration of DMSO. Worm lifespan assays were performed at 20° C.Animals that crawled off the plate or had an “exploded vulva” phenotypewere censored.

sir-2.1: sir-2.1 encodes the mammalian homolog of the NAD-dependentdeacetylase SIRT1 in C. elegans. SIR-2.1/SIRT1 activity is mediated byNAD level and is well described to be involved in longevity regulationin yeast, C. elegans, and Drosophila in response to caloric restriction.

aak-2/AMPK: aak-2 encodes the mammalian homolog of the AMP-dependentkinase AMPK in C. elegans.

daf-16 encodes a FOXO transcription factor.

eat-2 encodes a protein involved in pharyngeal pumping.

daf-2 encodes for the insulin-like growth factor 1 (IGF-1) receptor inC. elegans.

Results are shown in Table 16 and FIGS. 6A-6F. As shown in the figure,urolithin A significantly extended lifespan in wild-type C. elegans by75%, showing that urolithin A is a pro-longevity compound.

TABLE 16 Lifespan Extension in sir-2.1, aak-2, daf-16, daf-2, and eat-2Mutant C. elegans Lifespan Urolithin/ Median extension P-values DMSOlifespan compared to against Death/Censored Genotype conc. (days)control (%) control (trials) wild-type 1% DMSO 16 +75 <10⁻³ 37/24 (1)(N2) 50 μM UA 28 50/10 (1) sir-2.1 1% DMSO 21 +9.5 NS 46/14 (1) (mu86)50 μM UA 23 39/21 (1) aak-2 1% DMSO 16 +12.5 0.003  55/5 (1) (o524) 50μM UA 18 43/17 (1) daf-16 1% DMSO 16 +43.7 <10⁻³ 42/18 (1) 50 μM UA 2349/11 (1) daf-2 1% DMSO 42 +14.2 NS 21/19 (1) 50 μM UA 48 39/21 (1)eat-2 1% DMSO 21 +76.2 <10⁻³ 40/20 (1) 50 μM UA 37 26/34 (1)

Energy Stress—Caloric Restriction Pathway

The sir-2.1 mutant completely suppressed the activity of urolithin A onthe extension of median lifespan in C. elegans. This indicates thatsir-2.1 is required for the lifespan extension induced by urolithin Aand that urolithin A is dependent on sir-2.1 to extend lifespan.

The aak-2 mutant significantly suppressed the lifespan extensionphenotype induced by urolithin A from a 75% increase in mean lifespandown to a 12.5% extension. This indicates that aak-2 plays an importantrole in the lifespan subsequent to urolithin A exposure. This alsodemonstrates that urolithin A activation of AMPK is a key step in thelifespan extension induced by urolithin A.

Insulin Signaling Pathway

The daf-2 mutant completely suppressed the activity of urolithin A onthe extension of median lifespan in C. elegans. This indicates thatdaf-2 is required for the lifespan extension induced by urolithin A andthat urolithin A is dependent on daf-2 to extend lifespan.

Treatment of C. elegans with either rapamycin, resveratrol, or metformininduces an increase in worm mean lifespan, which has been shown to be aresult of acting on the autophagy pathway.

It is worth noting that the lifespan extension induced by urolithin Atreatment (75%) is much greater than the life extension induced byeither resveratrol (12.6%), rapamycin (19%), or metformin (27%) alone.

Example 7

Urolithin A Increases Mitochondrial Activity in C. elegans

C. elegans strains were cultured at 20° C. on nematode growth media(NGM) agar plates seeded with HT115 bacteria and containing 50 μMurolithin A or a corresponding concentration of DMSO as a control. Theworms were treated from eggs to the first day of adulthood. The strainsused were the SJ4103 (zcIs14[myo-3::GFP(mit)]), which is a stabletransgenic line expressing a mitochondrially localized green fluorescentprotein (GFP) with a cleavable mitochondrial import signal peptide underthe control of the specific body wall muscle promoter myo-3. GFPexpression and quantification was carried out according to the protocolpreviously described. Durieux J et al. (2011) Cell 144:79-91. Worms weretreated with 50 μM urolithin A from eggs and GFP was monitored after oneday of adulthood. Fluorimetric assays were performed using a Victor X4multilabel plate reader (Perkin-Elmer Life Science). Eighty worms werepicked at random (20 worms per well of a black-walled 96-well plate),and each well was read four times and averaged.

The results in FIG. 7 show that treatment of worms with urolithin Ainduces the expression of the mitochondrial GFP reporter driven by themuscle-specific myo-3 promoter in C. elegans. This striking increase inGFP expression provides clear evidence that mitochondrial capacity wasincreased due to the urolithin A. To permit such an increase in observedGFP signal, mitochondria in muscle must either be enlarged or morenumerous in these worms.

Example 8

Urolithin A Increases Mitochondrial Function in Aged C. elegans

C. elegans strains were cultured at 20° C. on nematode growth media(NGM) agar plates seeded with E. coli strain OP50. Strain used waswild-type Bristol N2 provided by the Caenorhabditis Genetics Center(University of Minnesota). Urolithin A was dissolved in DMSO. Animalswere exposed to compounds from eggs on plates seeded with live OP50bacteria. Control plates were prepared with the correspondingconcentration of DMSO (0.1%).

Measurement of oxygen consumption is a direct indicator of mitochondrialactivity. The effect of urolithin A on mitochondrial activity in aged C.elegans (10 days old) was assessed by treating C. elegans with urolithinA for 10 days of adulthood, at which time oxygen consumption wasmeasured using the Seahorse XF24 equipment (Seahorse Bioscience Inc.,North Billerica, Mass.). 250 ten-day-old C. elegans were used percondition. C. elegans were recovered from NGM plates with M9 medium,washed three times in 2 mL M9 to eliminate residual bacteria, andresuspended in 500 μL M9 medium. Worms were transferred into 24-wellstandard Seahorse plates (#100777-004) (50 worms per well) and oxygenconsumption was measured. The basal oxygen consumption of the worms wasfirst measured over 30 minutes at 5-minute intervals (0 min, 5 min, 15min, 20 min, 25 min, and 30 min) with 5 replicates per interval.Respiration rates were normalized to the exact number of worms per welldetermined after the completion of the experiment using astereomicroscope. After determining the basal oxygen consumption,uncoupled oxygen consumption was measured by addingcarbonylcyanide-p-(trifluoromethoxy) phenylhydrazone (FCCP) at the 30minute time point to the media in order assess the maximal oxygenconsumption capacity and maximal mitochondrial capacity. Uncoupledoxygen consumption was measured at 5-minute intervals (35 min, 40 min,45 min, 50 min, 55 min, and 60 min) to permit measuring themitochondrial function over time.

FCCP is a chemical uncoupling agent that abolishes the obligatorylinkage between the respiratory chain and the phosphorylation systemwhich is observed with intact mitochondria. This effect is due to theamphipathic properties of the molecule which dissolves in mitochondrialphospholipid bilayers. This dramatically increases ionic permeability ofthe mitochondrial membrane and generates dramatic proton leak leading toincrease in oxygen consumption due to the quenching by oxygen of theelectrons pumped into the respiratory chain in parallel to the protonleak. Since this oxygen consumption is dissociated (uncoupled) to ATPproduction (oxidative phosphorylation), FCCP increases oxygenconsumption while decreasing the generation of energy (ATP) by themitochondria. Fully uncoupled mitochondria, as achieved with FCCP,display the maximal capacity of their mitochondrial respiratory chain(maximal oxygen consumption) without the “brake” that oxidativephosphorylation and energy production represents.

The results depicted in FIGS. 8A and 8B illustrate that urolithin Aincreases the maximal mitochondrial capacity of 10-day-old aged C.elegans, as depicted by a prolonged effect on increased uncoupledrespiration in worms treated with urolithin A versus control(DMSO)-treated worms. Control untreated worms showed a brief increase inuncoupled respiration which quickly returned to basal levels of oxygenconsumption. Urolithin A-treated worms showed a more extended elevationin oxygen consumption. The extent of enhanced mitochondrial activity isshown by comparing the area under the curves (AUC) during the decouplingperiod with the average coupled respiration employed as the baseline. Itwas observed that urolithin A significantly increased uncoupledrespiration in aged worms as compared to control untreated worms overthe 30-minute period evaluated. FIG. 8C shows that aged, 10-day-old C.elegans have a decrease in basal respiration in comparison with young,1-day-old C. elegans. FIG. 8D shows that treatment of worms withurolithin A at 30 μM normalizes the respiration rates, and aged,10-day-old C. elegans show comparable levels of oxygen consumption toyoung, 1-day-old C. elegans.

Example 9

Urolithin A Increases Autophagy in C. elegans

C. elegans strains were cultured at 20° C. on nematode growth media(NGM) agar plates seeded with OP50 bacteria and containing 50 μMurolithin A or a corresponding concentration of DMSO as a control. Theworms were treated from eggs to the second day of the adulthood. Thestrains used were the DA2122 (adIs2122[lgg-1::GFP+rol-6(su1006)]), whichis a transgenic line expressing the LGG-1 protein (homolog of LC3 inhuman) fused with the green fluorescent protein (GFP). For pictureacquisition, worms were immobilized with tetramisole (Sigma) and mountedon 6% agarose pads on glass slides. Images were acquired using Zeiss LSM700 upright confocal microscope (Carl Zeiss AG, Oberkochen, Germany)from the same part of C. elegans. For each condition multiple worms wereobserved and imaged with the same initial parameters. Image processingand quantification of autophagic events was performed with the Fijisoftware.

The results depicted in FIGS. 9A and 9B illustrate that urolithin Aincreases the number of autophagic events, as depicted by the increasednumber of LGG-1::GFP dots per focal field (plan) in worms treated withurolithin A versus control (DMSO)-treated worms. This observation wasconfirmed by the fact that food deprivation, a dietary intervention thatincreases autophagy in C. elegans, shows the same profile as urolithinA-treated worms. This result provides clear evidence that urolithin Atreatment is an inducer of autophagy in worms and could explain thelongevity phenotype associated with such a treatment, as it has beenalready described that induction of autophagy, by dietary manipulationor treatment with specific compounds, increases life expectancy in anumber of animal species. Hansen M et al. (2008) PLoS Genetics 4:e24;Eisenberg T et al. (2009) Nat Cell Biol 11:1305-14; Bjedov I et al.(2010) Cell Metab 11:35-46.

Example 10

The Autophagic Pathway is Involved in the Longevity Phenotype Induced byUrolithin A in C. elegans

Lifespan tests were performed as described. Mouchiroud L et al. (2011)Aging Cell 10:39-54. Briefly, worms were observed using a Nikon SMZ1500stereomicroscope (Nikon, Melville, N.Y., USA). Briefly, 10 L4 worms weretransferred on plates containing NGM medium with vehicle (DMSO 0.1%,final concentration) or urolithin A dissolved in DMSO (finalconcentration 50 μM) and seeded with E. coli RNAi clones as indicated.After 3 days (corresponding to day 0), newly developed 60-100 L4 wormswere used per condition, scored and transferred to fresh plates every 3days starting from day 1 (young adult worms). All lifespan experimentswere performed at 20° C. Animals that crawled off the plate or had an“exploded vulva” phenotype were censored.

Survival analyses were performed using the Kaplan Meier method and thesignificance of differences between survival curves calculated using thelog rank test. Differences between two groups were assessed usingtwo-tailed t-tests. Analysis of variance, assessed by Bonferroni'smultiple comparison test, was used when comparing more than two groups.The statistical software used was GraphPad Prism 5 (GraphPad Software,Inc.) and all p-values<0.05 were considered significant.

Gene silencing was carried out as previously described by using the RNAinterference (RNAi) process. Mouchiroud L et al. (2011) Aging Cell10:39-54. In C. elegans, RNAi is made by feeding. Briefly, the nematodeswere fed with bacteria modified with a plasmid encoding a dsRNAtargeting a gene of interest. The worms eat these bacteria which willsimply allow the inactivation of this gene. As a control, worms were fedwith the RNAi clones HT115, which encodes an empty vector.

To determine the requirement of the autophagic pathway for the lifespanphenotype induced by urolithin A treatment (FIGS. 10A-10C), two genesencoding major autophagic proteins were inhibited by RNA interference(RNAi). These two proteins were vps-34 (sequence name: B0025.1, homologof VPS34 in human), which encodes a phosphatidylinositol 3-kinase (PI3K)that regulates multiple steps in vesicular trafficking and that isrequired for autophagy mechanism, and bec-1 (sequence name: T19E7.3,homolog of beclin in human), which encodes a class IIIphosphatidylinositol 3-kinase complex that plays a role in localizingautophagy proteins to preautophagosomal structures. The inhibition oftheir expression by RNAi has already been described to suppress thelifespan phenotype induced by dietary restriction, which is anexperimental condition that promotes autophagy in C. elegans. Hansen Met al. (2008) PLoS Genetics 4:e24.

TABLE 17 Proteins Involved in the Autophagy Pathway in Different SpeciesYeast C. elegans Mouse Name Name Name Role in Autophagy ATG8 LGG-1 LC3Autophago some formation-marker of autophagosome membrane ATG6 BEC-1BECN1 Beclin1 regulates the kinase activity of Vps34 at the endoplasmicreticulum Vps34p VPS-34 VPS34 Vps34 has been shown to interact withVps15, a protein kinase. Vps15 can activate the lipid kinase activity ofVps34 and interact with Rab5, which has been hypothesized to recruit theVps34/15 complex to early endosomes.

The results depicted in FIGS. 10A-10C illustrate that urolithin Aincreased the lifespan of worms through the activation of the autophagymechanism. The fact that the lifespan extension observed in wormstreated with urolithin A and fed with the empty vector is suppressedwhen these worms are fed with bacteria expressing RNAi against vps-34 orbec-1 reveals that these two proteins are each required and essentialfor the longevity phenotype. Such an observation shows that urolithin Atreatment promotes the activity of vps-34 and bec-1, and leads to theinduction of the autophagy mechanism to extend longevity.

Example 11

Urolithins Maintain Muscle Function in C. elegans During the AgingProcess

To examine the effect of urolithins on muscle function, pharyngealpumping was examined in C. elegans worms at day 7 and day 14. Worms wereobserved using a Nikon SMZ1500 stereomicroscope (Nikon, Melville, N.Y.,USA). Ten L4 worms were transferred on plates containing NGM medium withvehicle (DMSO 0.1%, final concentration) or the compound of interest(urolithin A (UA), urolithin B (UB), urolithin C (UC), urolithin D (UD))dissolved in DMSO (final concentration 50 μM) and seeded with E. coliRNAi clones as indicated. After 3 days (corresponding to day 0), newlydeveloped 60 to 100 L4 worms were used per condition, scored andtransferred to fresh plates every 3 days starting from day 1 (youngadult worms). Experiments were performed at 20° C. Animals that crawledoff the plate or had an “exploded vulva” phenotype were censored.

The pharyngeal pumping rates at day 7 and 14 were measured for 10 wormsby transferring single worms to an unseeded plate and scoring forpharyngeal pumping under a dissecting microscope for 30 seconds. Thepharyngeal pumping rates were then computed by extrapolating the pumpingmade by worms during this 30 second period.

Pharyngeal pumping at day 7 were markedly increased for all urolithinstested (FIG. 11 ), demonstrated the ability of urolithin treatment toimprove muscle function in young worms. This improved muscle function ismaintained in aging worms and urolithin treatment also improves musclefunction in worms at day 14. In the aged worms (day 14) urolithintreatment, and in particular urolithins A and B, decreases the agerelated decline in muscle activity (pharyngeal pumping).

To examine the effect of urolithins on muscle activity during aging, C.elegans mobility was examined over time. Synchronized populations ofwild-type nematodes were prepared by standard methods and cultivated at20° C. on NGM agar containing E. Coli (0P50) with or without testcompounds (ellagic acid (EA), urolithins A (UA), and B (UB)). At Day 0,five plates were prepared, each containing 10 young adult worms. Wormswere assayed for mobility at different ages, day 1, 3, 5 and 8 ofadulthood. The wild type (N2, Bristol) worm strain was employed forthese studies.

For C. elegans movement tracking, 45 seconds of video were recordedusing a Nikon DS-L2/DS-Fi1 camera and controller setup, attached to botha computer and a standard bright field microscope, tracking an entireplate of 10 worms, with 5 plates per condition. The movement of wormsduring this time was calculated using an adapted version of thefreely-available software Parallel Worm Tracker for MATLAB. The totalmovement over the 45 second tracking was averaged across all worms foreach condition. This experiment was repeated twice with between 40 and70 worms per condition.

Ellagic acid treatment has no effect on this age related mobilitydecline (FIG. 12 ). In contrast, urolithin treatment with 50 μM ofeither urolithin A or B, demonstrate a marked ability to reduce musclefunction decline and maintain muscle function in aging C. elegans worms.This demonstrates the ability of urolithin treatment to preserve musclefunction during the aging process.

To further characterize the effect of urolithins on muscle function andmobility, C. elegans were treated and their mobility observed undertime-lapse microscopy. The effects of urolithin A on muscle function inC. elegans were examined. Briefly, worms were observed using a NikonSMZ1500 stereomicroscope (Nikon, Melville, N.Y., USA). Briefly, 5 wormscoming from different plates were transferred on plates containing NGMmedium with vehicle (DMSO 0.1%, final concentration) or compound ofinterest (ellagic acid (EA), urolithin A (UA), urolithin B (UB),urolithin C (UC), urolithin D (UD)) dissolved in DMSO (finalconcentration 50 μM) and seeded with E. coli RNAi clones as indicated.Time lapse was performed by recording 20 pictures of mobile worms with a10 second interval by using a Zeiss Axioplan-2 microscope (Carl ZeissMicroImaging, Thornwood, N.Y., USA). Worm tracking was realized usingthe ImageJ software. Experiments were repeated three times.

Urolithin treatment resulted in a marked increase in motility that wasmaintained to varying degrees during the aging process (FIG. 13 ). Thestrongest effect was observed for urolithin B. The results presentedhere demonstrate the ability of all urolithin treatments to improvemuscle function in C. elegans and preserve muscle function against agerelated muscle activity decline.

Example 12 Urolithins Induce Autophagy in Mammalian Cells

The ability of urolithins to induce autophagy in mammalian cells wasalso examined. ModeK cells derived from intestine, were cultured in RPMI1640 medium including 4.5 g/L glucose, 10% fetal calf serum, 1 mM SodiumPyruvate, 10 mM HEPES, 0.1 mM NEAA, 2-Mercaptoethanol Penicillin 100UI/mL and Streptomycin 100 μg/mL. Cells were cultured at 37° C. under a5% CO2 atmosphere. Cells were treated with increasing concentrations ofurolithin A (10 μM, 20 μM and 50 μM). Treatment was performed for eighthours, after which cells were lysed and prepared for Western Blotanalysis. Protein expression levels were examined for autophagy relatedproteins LC3-I and II (Cell Signaling antibody #4108), p62 (CellSignalling antibody #5114), AMPKα (Cell Signaling antibody #2603), andp-AMPKα (Cell Signaling antibody #2531). The housekeeping protein, HSP90(BD Transduction Laboratories antibody #610418) level was measured as aloading control.

Increasing concentrations of urolithin A resulted in increasing levelsof phosphorylated AMPKα (FIG. 14 ). The ratio of p-AMPKα/AMPKα increasedin a dose-dependent manner in response to treatment with urolithin A,ranging from 4-fold to over 10-fold induction. Urolithin A treatmentalso resulted in significant increases in the LC3-II/LC3-I ratio (ahallmark of autophagy), at all doses tested. The ratio of LC3-II/LC3-Iincreased as high as 10-fold in treated cells as compared to thevehicle-treated cells at the doses tested. The levels of p62 decreasedwith the addition of urolithin A, in accordance with autophagy process.These results demonstrate the ability of urolithin A to induceautophagy.

Induction of autophagy by urolithins was also demonstrated in mammalianprimary hepatocytes. Primary hepatocytes were isolated by perfusionthrough the supra-hepatic inferior vena cava from 7- to 9-week-oldwildtype C57BL/6J mice, as described previously (Ryu et al., 2011).Isolated primary hepatocytes were cultured in Medium 199 including 4.5g/L glucose, 10% fetal calf serum, 0.1 mM NEAA, 10 mM HEPES and 50 μg/mLgentamicin. These cells were then treated with either 10 μM, 20 μM or 50μM of urolithin A for a 16 hour period, after which cells were lysed andprepared for Western Blot analysis. Protein expression levels wereexamined for autophagy related proteins LC3-I and II (Cell Signalingantibody #4108), p62 (Cell Signalling antibody #5114), AMPKα (CellSignaling antibody #2603), and p-AMPKα (Cell Signaling antibody #2531).The housekeeping protein, HSP90 (BD Transduction Laboratories antibody#610418) level was measured as a loading control.

Urolithin A treatment at 20 and 50 μM led to increased levels ofphosphorylated AMPKα, with in treated cells the ratio of p-AMPKα/AMPK-αsignificantly rising well above 10-fold the ratio of p-AMPKα/AMPK-αobserved in control cells. Importantly, a dose-dependent effect andincrease in the ratio of LC3-II to LC3-I was observed followingtreatment with urolithin A, a hallmark of autophagy (FIG. 15 ). P62protein levels were observed to decrease following exposure to urolithinA. This dose effect of urolithin A on the ratio of LC3-II/LC3-I proteinratio, along with the decrease in p62, demonstrates the ability ofurolithin A to induce autophagy in mammalian primary hepatocytes anddemonstrates a general autophagy activity of urolithins in primarycells.

Example 13 Urolithins Induce Autophagy Across Species in Mammalian Cells

The ability of urolithins to induce autophagy in muscle cells acrossspecies was examined using mouse C2Cl2 myoblasts and human primaryskeletal muscle cells.

C2Cl2 myoblast were cultured in Dulbecco's modified Eagle's medium(DMEM) including 4.5 g/L glucose, 20% fetal calf serum, and 50 μg/mLgentamicin. Urolithin A was dissolved in DMSO in a stock solution of 1mM. Cells were treated at final concentrations of urolithin A of 10 μM,20 μM, and 50 μM for a period 16 hours. Control cells were treated withDMSO at an equivalent final concentration for the same period and servedas the untreated control.

Human primary skeletal myocytes were cultured in vitro and exposed toincreasing concentrations of urolithin A, including 10 μM, 20 μM, and 50μM for 16 hours. Human skeletal myoblasts were grown in DMEM plus 2%horse serum. Control cells were treated with DMSO at an equivalent finalconcentration for the same period and served as the untreated control.

At the end of treatment, cells were lysed with RIPA buffer and appliedto SDS-PAGE and analyzed by western blot. Protein expression levels wereexamined for autophagy-related proteins LC3-I and LC3-II (Cell Signalingantibody #4108), p62 (Cell Signaling antibody #5114), AMPKα (CellSignaling antibody #2603), and p-AMPKα (Cell Signaling antibody #2531).The housekeeping protein, β-actin (Cell Signaling Antibody #4967) levelwas measured as a loading control. Representative results are shown inFIGS. 16 and 17 .

In both mouse C2Cl2 muscle cells (FIG. 16 ) and human primary myoblasts(FIG. 17 ) urolithin A treatment led to a dose-dependent increase inautophagy as verified by an increase in the ratio of LC3-II to LC3-I,accompanied by a decrease in the levels of p62, as observed by thechange in density of the bands in the western blots. Also,phosphorylated AMPKα (pAMPKα) levels increased, with the ratio ofpAMPKα/AMPKα increasing several fold in response to exposure tourolithin A.

Example 14 Urolithins Induce Autophagy in Primary Cells

Human Aortic Endothelial Cells (HAOEC) are primary endothelial cellsisolated from normal human aorta. They were sourced from CellApplications Inc. (CAI), San Diego, USA and obtained cryopreserved atthe second passage from the European Collection of Cell Cultures(ECACC). The cells were thawed and cultured with Endothelial Cell GrowthMedium, provided by the ECACC, catalog number 06091509 (CAI No.211-500), in a 37° C., 5% CO₂ humidified incubator; media was changed 24hours after thawing. Endothelial Cell Growth Medium was changed everyother day until the cells reached 80% confluency. Cells were thensubcultured into four 6-well dishes and after overnight incubation weretreated with different concentrations of urolithin A, 10 μM, 20 μM, and50 μM, or a DMSO control for a period of 16 hours, 3 wells pertreatment. Control cells were treated with DMSO at an equivalent finalconcentration for the same period and served as the untreated control.

At the end of treatment, cells were lysed with RIPA buffer and appliedto SDS-PAGE and analyzed by western blot. Protein expression levels wereexamined for autophagy-related proteins LC3-I and LC3-II (Cell Signalingantibody #4108), and p62 (Cell Signaling antibody #5114). Thehousekeeping protein, HSP90 (BD Transduction Laboratories antibody#610418) level was measured as a loading control. Representative resultsare shown in FIG. 18 .

In human primary aortic endothelial cells (FIG. 18 ) urolithin Atreatment led to a dose-dependent increase in autophagy as verified byan increase in the ratio of LC3-II to LC3-I to as high as 3-foldinduction at 50 μM of urolithin A, which was also accompanied by adecrease in the levels of p62, as observed by the change in density ofthe bands in the western blots. These results clearly indicate thaturolithins increase autophagy in endothelial cells.

Example 15 Urolithins Induce Autophagy in Mammals

To determine the ability of urolithin A to induce autophagy in vivo,healthy 10-week-old C57BL/6J mice were treated with either a standardrodent diet or a diet containing urolithin A mixed with food to reach adosing of 55 mg/kg/day delivered to the mice. Following 8 weeks oftreatment with urolithin A, mice were sacrificed and the liver of eachanimal was surgically isolated. The isolated livers were then placed inlysis buffer and prepared for western blot analysis.

Protein expression levels were examined for autophagy related proteinsLC3-I and LC3-II (Cell Signaling antibody #4108), p62 (Cell Signallingantibody #5114), AMPKα (Cell Signaling antibody #2603), and p-AMPKα(Cell Signaling antibody #2531). The housekeeping protein, HSP90 (BDTransduction Laboratories antibody #610418) level was measured as aloading control. Each lane in the Western blot represents a proteinsample for the liver of a single mouse, to permit observation of theeffects aver a number of treated animals.

Urolithin A treatment at an oral dose of 55 mg/kg/d that was admixedinto the animal chow diet led to increased levels of phosphorylatedAMPKα (pAMPKα), as well as an increase in the ratio of LC3-II to LC3-I,a hallmark of autophagy, in a healthy mouse liver following oralconsumption (FIG. 19 ). The increase in autophagy induced by the oraltreatment was observed to be on average approximately 400% over thecontrol untreated mice based on the ratio of LC3-II/LC3-I. p62 proteinlevels also decreased with utolithin A treatment. These resultsdemonstrate the ability of urolithin A to induce autophagy in mammaliancells and tissues following oral consumption.

Example 16 Urolithin A Improves Motor Activity in Mammals

To examine the effect of urolithin A treatment on activity and musclefunction in mammals, 10-week-old C57BL/6J mice were treated with eithera standard rodent diet or a diet containing urolithin A mixed with foodto reach a dosing of 55 mg/kg/day delivered to the mice. Following 11weeks of treatment with urolithin A, a 23 cm in diameter running wheelwas placed in the cage of each mouse. Spontaneous exercise activity wasmeasured over a five day period at 20 min intervals, allowing thecapture of the circadian rhythm of the exercise activity.

Urolithin A treatment resulted in a significant and sustained increasein the distance covered by the mice each day (FIG. 20 ). During theinitial 3 days the urolithin A treated animals showed an increase of 25%over untreated controls in the cumulative distance covered. Thisdifference increased further to 29% on the final two days tested. Theseresults demonstrate the ability of urolithin A treatment to improvemotor activity in mammals. A clear training effect was observed in whichmice administered urolithin A not only increased their spontaneousrunning activity, but there was an increase over time. Additionally,this demonstrated that the benefits persist over time.

Example 17 Urolithin A Improves Motor Activity in Aged Mammals

To examine the effects of urolithin A treatment on motor activity inaged mammals 17-month-old C57BL/6J mice were treated with either a highfat rodent diet or a high fat rodent diet containing urolithin A mixedwith food to reach a dosing of 50 mg/kg/day delivered to the mice.

Following 30 weeks of treatment with urolithin A, mice were housedindividually and a 23 cm diameter running wheel was placed in the cageof each mouse. Spontaneous exercise activity was measured continuouslyover a two day period allowing the capture of the circadian rhythm ofthe exercise activity. Afterwards, mice were returned to their homecages.

Mice became habituated to the presence of the activity wheel on day 1.To measure the effect of urolithin A treatment on activity as well asmuscle function, the cumulative distance that animals covered during thepeak running period (dark phase) of day 2 was measured. As shown in FIG.21 , urolithin A treatment resulted in an increase of 57% (p<0.05) incumulative distance run in treated aged mice fed a high fat diet. Thisclearly shows the positive benefits of urolithins on spontaneousactivity and running during aging. This provides evidence that thedelivery of urolithin to an aging population will improve locomotoractivity and running ability and help prevent the natural declineobserved.

Example 18 Urolithin A Improves Muscle Strength in Aged Mammals

To examine the effects of urolithin A treatment on muscle function inaged mammals, 17-month-old C57BL/6J mice were treated with either a highfat rodent diet or a high fat rodent diet containing urolithin A mixedwith food to reach a dosing of 50 mg/kg/day delivered to the mice.

Following 26 weeks of treatment with urolithin A, mice were assessed fortheir neuromuscular function by means of a grip test. Using the griptest, one can measure the forelimb grip strength of mice. Mice wereplaced on a grid to enable their fore paws grasp the support. To measurethe grip strength, mice are pulled backward in the sensor axis untilthey are no longer able to hold the grid.

Representative results are presented in FIG. 22 . Mice receivingurolithin A showed a greater grip strength than untreated mice.

Example 19 Urolithin A Improves Locomotor Activity in Aged Mammals

Locomotor activity may be also assessed by measuring animal movementsusing infrared detection. To determine the effects of urolithin A onspontaneous locomotor activity in aged animals, 16-month-old C57BL/6Jmice were treated with either a standard rodent diet or a dietcontaining urolithin A mixed with food to reach a dosing of 50 mg/kg/daydelivered to mice. Following 34 weeks of treatment with urolithin A,mice were placed in a locomotor activity monitoring system, TSE Systems.Infrared transmitters and receivers are placed in the x, y, and z axis.Interruption of the infrared beams in the x, y axis by the passage ofthe mouse, allows for a measurement of the horizontal movement(ambulation). Interruption of the infrared beams in the z axis by themouse, allows for the measurement of rearing behavior. Each beaminterruption results in one count being recorded. Total counts weremeasured for each 30 minute period over a two day period. The totalcounts observed during the 12 hour dark period, which corresponds to thetime of highest activity in mice, were summed for each animal.

FIG. 23 shows that aged mice treated with urolithin A showed a 14%increase in ambulation and a 25% increase in rearing. These resultsdemonstrate the ability of urolithin A to increase mobility in agedanimals.

Example 20 Urolithin A Increases Autophagy in Skeletal Muscle of AgedMammals

Following 34 weeks of treatment, the mice from Example 19 weresacrificed and the organs were collected. The gastrocnemius muscles wereassessed for the presence of autophagy by the assessment of the changein the ratio of the autophagy markers LC3 and p62 using western blotanalysis. Each lane corresponds to a muscle sample from an individualmouse. Representative results are shown in FIG. 24 . The ratio ofLC3-II/LC3-I increased in the muscles of urolithin A (UA)-treated miceversus untreated mice. This was accompanied by a decrease in the levelsof p62 in the muscles of UA treated versus untreated mice. These shiftsin the levels of LC3 and p62 in the muscle of mice are consistent withan increase in the level of autophagy taking place. This demonstratesthat the oral administration of urolithin A leads to autophagy at theorgan level, including muscle. Mice receiving UA in their diets alsodisplayed an increase activation of AMPK in their muscle as observed byan increase in the ratio of p-AMPKα/AMPKα.

Example 21 Urolithins Induce Autophagy in Mouse Muscle Cells

To determine the ability of urolithin A to induce autophagy in mousemuscle cells, undifferentiated C2Cl2 mouse myoblasts were seeded in T25flasks and incubated overnight. Cells were incubated in κ% CO2 at 37° C.These cells were treated with one of the urolithins-urolithin A (UA),urolithin B (UB), urolithin C (UC) or urolithin D (UD)—dissolved in a0.1% solution of DMSO for 24 hours. In one experiment UA, UB, UC and UDwere dosed at a concentration of 100 μM for C2Cl2 myoblast treatment. Ina separate study, C2Cl2 myoblasts were incubated at increasingconcentrations of UA (10 μM, 50 μM and 100 μM). For each experiment acontrol culture was treated with 0.1% DMSO for the same period andserved as the untreated control. In the final 30 minutes of incubation,lysosome degradation inhibitor was added to the culture. The cells werethen washed with phosphate buffer solution and then treated with a 1%solution of trypsin enzyme to remove the adhesive cells from the cellculture dishes. The cells were then treated with EMD Millipore selectivepermeabilization reagent to permeabilize the cells so as to make theinternal cellular proteins accessible to antibody treatment. Cells werethen incubated with antibody to LC3B according to kit protocol (EMDMillipore FlowCellect™ Autophagy LC3 Antibody-based Assay Kit (Cat no.FCCH100171)). The cells were then washed and resuspended in the kitassay buffer. The resulting cells were then run through a flow cytometerand LC3B-FITC fluorescence was quantified to measure the level of LC3Bin C2Cl2 cells for the various treatments. When cells undergo autophagytheir intracellular levels of LC3-II increase which is detected by thismethod. This autophagy can be detected by fluorescent antibody labelingof LC3B, which can be measured quantitatively using flow cytometry. Anincrease in cellular autophagy is reflected by an increasing signal inthe histogram of LC3B FITC.

Urolithin A treatment of C2Cl2 cells results in an increase in LC3Bexpression that is dose dependent (FIG. 25 ) demonstrating the abilityof urolithin A to induce autophagy in mouse muscle cells in adose-dependent manner (10 μM, 50 μM and 100 μM). In a separate study,induction of autophagy was also observed in C2Cl2 mouse myoblastsfollowing treatment with urolithin A (UA), urolithin B (UB), urolithin C(UC) and urolithin D (UD) at 100 μM after 24 hrs of treatment (FIG. 26). Incubation with UA, UB, UC and UD all led to a increasing signal ofLC3B as shown by the positive shift of the histogram. This datademonstrates the ability of the urolithins as a class of compounds toinduce autophagy in mammalian myoblasts.

Example 22 Synthesis of Urolithins and Urolithin Precursors

Examples of urolithin compounds of the invention are shown in FIG. 27 ascompounds 1 through 16. Examples of urolithin precursors of thisinvention are shown in FIG. 27 as compounds 17 through 25.

FIG. 28 describes methods to synthesize the compounds 1 to 25 to enabletheir therapeutic use in the present invention. Several chemicalreactions are conserved and the details are provided below as ReactionsA, B, C and D.

Reaction A

Oxalyl chloride (1 eq.) and dimethylformamide (cat.) were added to asuspension of the benzoic acid (1 eq.) in dichloromethane. The reactionmixture was stirred for 16 hours at room temperature and concentratedunder reduced pressure. The residue was immediately solubilized indichloromethane and cooled to 0° C. The phenol (1.1 eq.) andtriethylamine (1.5 eq.) were successively added and the mixture wasstirred for 5 minutes. Acetonitrile was finally added and the resultingsolution was stirred for 16 hours at room temperature. The reaction wasquenched by addition of a saturated solution of ammonium chloride andthe layers were separated. The aqueous phase was extracted withdichloromethane and the combined organic layers were dried over sodiumsulphate, filtered and concentrated under reduced pressure. The crudewas purified by column chromatography on silica gel.

Reaction B

Sodium acetate (2 eq.), (S)-Phos (0.1 eq.) and palladium diacetate (0.1eq.) were successively added to a solution of the ester (1 eq.) indimethylacetamide under an argon atmosphere. The reaction mixture wasstirred at 130° C. for 3 days. The reaction was quenched by addition ofwater and the mixture was extracted with dichloromethane. The combinedorganic layers were washed with brine, dried over sodium sulfate andconcentrated under reduced pressure. The remaining dimethylacetamide waseliminated using a Hickman apparatus. The residue was purified by columnchromatography on silica gel.

Reaction C

Boron tribromide (1.25 eq. by methoxy moiety) was added to a solution ofthe protected urolithin (1 eq.) in chloroform. The mixture was thenheated up to 60° C. and stirred for 4 days. The reaction was quenched byaddition of methanol and the mixture was evaporated to dryness. Theresidue was purified by column chromatography on C18.

Reaction D

A mixture of the diphenol (2 eq.), the bromobenzoic acid (1 eq.) andNaOH (2.2 eq.) in water was heated at reflux and stirred for 2 hours.Water and CuSO₄·5H₂O (0.1 eq.) were added and the mixture was stirredunder reflux for an additional 1 hour. The reaction was then cooled downto room temperature and the precipitate was filtered on a glass frit(porosity 4). The product was then dissolved in absolute ethanol andconcentrated in order to remove water. The residue was dissolved in hotmethanol and filtered on paper.

1-113. (canceled)
 114. A composition, comprising a urolithin or prodrugor pharmaceutically acceptable salt thereof; and a second ingredientselected from the group consisting of resveratrol and coenzyme Q10;wherein the coenzyme Q10 is ubiquinol or ubiquinone.
 115. Thecomposition of claim 114, wherein the second ingredient is resveratrol.116. The composition of claim 114, wherein the second ingredient iscoenzyme Q10; and the coenzyme Q10 is ubiquinol.
 117. The composition ofclaim 114, wherein the second ingredient is coenzyme Q10; and thecoenzyme Q10 is ubiquinone.
 118. The composition of any one of claim114, wherein the urolithin is urolithin A.
 119. The composition of 114,further comprising a pharmaceutically acceptable carrier.
 120. Acomposition, comprising a urolithin or a prodrug or pharmaceuticallyacceptable salt thereof; resveratrol; and coenzyme Q10; wherein thecoenzyme Q10 is ubiquinol or ubiquinone.
 121. The composition of claim120, wherein the coenzyme Q10 is ubiquinol.
 122. The composition ofclaim 120, wherein the coenzyme Q10 is ubiquinone.
 123. The compositionof claim 120, wherein the urolithin is urolithin A.
 124. The compositionof claim 120, further comprising a pharmaceutically acceptable carrier.125. A method of improving or increasing autophagy, comprisingadministering to a vertebrate in need thereof an effective amount of thecomposition of claim
 114. 126. The method of claim 125, wherein theautophagy is mitophagy.
 127. A method of treating, preventing, ormanaging a disease selected from the group consisting of muscledegenerative disorder, sarcopenia, muscular dystrophy, cardiomyopathy,Ulrich myopathy, obesity, cardiovascular disease, atherosclerosis, and amitochondria-related disease associated with altered mitochondrialfunction or a reduced mitochondrial density; or a disease related to ametabolic function; comprising administering to a vertebrate in needthereof an effective amount of the composition of claim
 114. 128. Amethod of preventing, treating, or managing a condition selected fromthe group consisting of frailty, muscular atrophy, muscle disuseatrophy, skeletal muscle atrophy or impaired muscle strength associatedwith aging, and skeletal muscle damage; a mitochondria-related conditionassociated with altered mitochondrial function or a reducedmitochondrial density; a condition related to a metabolic function; inprotecting against muscle damage or injury; in protecting or maintainingcardiac muscle; in improving muscle function, muscle strength, musclemass, muscle endurance, muscle recovery following exercise; in improvingbalance or coordination; in improving mitochondrial function; or inweight management; comprising administering to a vertebrate in needthereof an effective amount of the composition of claim
 114. 129. Amethod of treating a disease or condition selected from the groupconsisting of sarcopenia, sarcopenia of aging, osteoarthritis, muscularatrophy, muscle disuse atrophy, skeletal muscular atrophy, cardiacdeterioration, cardiac deterioration with aging, negative consequenceslinked to aging; improving activity during aging, reducing age-relatedcataracts, improving balance or coordination, and frailty, comprisingadministering to a subject in need thereof a therapeutically effectiveamount of the composition of claim
 114. 130. A method of improving orincreasing autophagy, comprising administering to a vertebrate in needthereof an effective amount of the composition of claim
 120. 131. Amethod of treating, preventing, or managing a disease selected from thegroup consisting of muscle degenerative disorder, sarcopenia, musculardystrophy, cardiomyopathy, Ulrich myopathy, obesity, cardiovasculardisease, atherosclerosis, and a mitochondria-related disease associatedwith altered mitochondrial function or a reduced mitochondrial density;or a disease related to a metabolic function; comprising administeringto a vertebrate in need thereof an effective amount of the compositionof the composition of claim
 120. 132. A method of preventing, treating,or managing a condition selected from the group consisting of frailty,muscular atrophy, muscle disuse atrophy, skeletal muscle atrophy orimpaired muscle strength associated with aging, and skeletal muscledamage; a mitochondria-related condition associated with alteredmitochondrial function or a reduced mitochondrial density; a conditionrelated to a metabolic function; in protecting against muscle damage orinjury; in protecting or maintaining cardiac muscle; in improving musclefunction, muscle strength, muscle mass, muscle endurance, musclerecovery following exercise; in improving balance or coordination; inimproving mitochondrial function; or in weight management; comprisingadministering to a vertebrate in need thereof an effective amount of thecomposition of claim
 120. 133. A method of treating a disease orcondition selected from the group consisting of sarcopenia, sarcopeniaof aging, osteoarthritis, muscular atrophy, muscle disuse atrophy,skeletal muscular atrophy, cardiac deterioration, cardiac deteriorationwith aging, negative consequences linked to aging; improving activityduring aging, reducing age-related cataracts, improving balance orcoordination, and frailty, comprising administering to a subject in needthereof a therapeutically effective amount of the composition of claim120.